Kind: captions
Language: en
alhamdulillah we can gather here
together this morning in such a
fortunate event of Friday morning
lecture on BI refiner already present
here with us is Professor ramaraj buatti
from Nicole State University USA to give
the talk and I'm very thankful for
Professor Raj as we actually have 13
hours
different yeah yeah so it is in the
evening in the US now
and I will first introduce Professor Raj
buatti he is the LC fourer distinguished
service professor and John Brady senior
and John Brady Junior and professor in
nicool State University Luciana Us in
Department of biological
science he did his Bachelor to master uh
to doctoral degree in India and go to
various part of the world before uh
settled in us and he is the editor of
many reputable
Journal Environmental Quality and
management cure and pollution report
applied science and I cannot mention all
of them because there are so
many and his field of Interest are
antibiotic resident uh resistance
microbiology bior remediation ethanol
production and
biodegradation and well there's a long
list of
achievement but last year he got uh he
was awarded worldclass Professor from
Indonesian
government that is the most important
for us because we are in
Indonesia and without further
delay uh we would like to welcome
Professor Raj to give the talk on lnos
cus bioethanol and microbial lipid for
bioenergy okay Professor Raj the time is
yours all right thanks Dr P it's good to
see you all and today this is my fourth
lecture in this lecture series and today
I'm going to talk about biofuels and
mostly
liosan um let me share my slides um get
my slides open yes yeah all right um so
what I'm going to talk about because of
the mixed audience students in different
um you know classifications from
graduate student to undergrad I'm going
to start with some basic um what is
ethanol what is cellulosic ethanol and
then talk about the current technology
in the US commercial level and then uh
complete my talk with my uh personal
research on this field uh how we going
to use this L material holistically not
only the sugar but also liin um using
microbial liquid as using liin to
convert to microbial liquid so I'm going
to start with basic and and finish with
applied research okay
um so first of all G Green Technology is
the best word now Green Technology is
continuously evolving group of
methods constantly developing people are
um you know finding new methods uh not
only generating energy to also various
processes including non-toxic cleaning
products uh the most urgent issue here
is the alternative fuels because the
petroleum is going to run out eventually
so you need to find other other fuel
sources especially the renewable ones so
uh Green Technology in the alternative
Fiel is used to be a really hot topic
but um it's catching up again after we
go through this o petroleum oil price
come down now we're getting again back
on the research so it's a if you look at
this Green Technology it encompasses a
lot of things it does the
sustainability uh it also does Source
reduction of waste material in
innovation of course there a lot of new
ideas new methods developed and also
socioeconomic aspect of it and viability
CR Cradle to cradle design so it it's
really a good deal it holistic approach
uh it combines a lot of things Green
Technology okay so let's talk about
ethanol so ethanol is this molecule C2
H5
um2 this one is the molecular model and
we have the since it has oxygen it is a
better burning fuel right so you
actually blend 10% ethanol in Us in all
the gasoline product
nowadays so if you look at the
fermentation reaction the starting
material is sugar uh mostly glucose and
then yeast convert glucose to ethanol
and carbon dioxide that's a simple
design um but you also get two molecules
of carbon dioxide from one molecule of
glucose and two molecular ethanol you
get and just to show you the structure
of how it goes through the py weight
through glycolysis and then you have uh
ethanol production through acetal deide
okay so just to show you the simple
schematic so from glucose through
glycolysis molecular to pyate and the
East gets two ATP for its energy source
and then pyate is converted to acetal
deide and then acetal ethanol and this
NAD is constantly recyle um going back
to generate more ATP so this is a
simplistic view of um the the
biochemistry in glucose ferment ation to
ethanol
um if you look at grain ethanol
it basically come from grain in US is
mostly corn and then other countries in
Brazil they use sugar cane and then
other grains as well and potatoes is
another source you get um anywhere any
any material that are starch you could
use it for this purpose okay so it's
very energy efficient is 25% more energy
than is used to grow Harvest and distill
into it so if you look at energy output
energy input
ratio it's it's it's positive side 1.6
is the ratio okay from output to
input if you compare gasoline with with
ethanol ethanol is very comparable so
Octan a number in E85 85% ethanol um
fuel is 100 compared to gasoline 86 to
90 for um the fuel source of gasoline is
crude oil but here you have variety of
agriculture product you could use and
BTU is also comparable with
70% of energy ratio in ethanol and both
are liquid so we need liquid fuel for
transportation purpose so it's really
good uh a little bit of more
biochemistry so the starting material is
a corn or potato any starch material so
you have to hydrolize and convert come
to sugar and then and glucose is through
glycolysis you convert yeast converted
to pyu and then use py DEC carboxilate
convert py to acet alide and then
alcohol
dehydrogenase convert acet alide to
ethanol at a simple biochemical pathway
and just to show what are the enzymes
involved you need to have convert star
to glucose you need mlas um and then
from glucose to pyade
the ATP is transferred and then you had
pyro for oxygen oxid reductase enzyme
convert pyro to acet co and AC acetogen
convert AC Co to AC
alide and
thenal then the enzy ethanol dehydrogen
convert them to ethanol so these are
various steps in fermentation
reaction so if you look at holistically
so most of the East use the um car six
carbon sugar um fermentation and we also
have pentos using sugar East like Pia
stas okay so we have uh the the process
of glycolysis and also simulation
process use them to generate energy
fermentation reaction then pentos
phosphate pathway so if you look at it
you have from hexo sugar transporter can
transport the sugar inside the yeast and
then from there it goes through um uh
glycolysis process and then if it you
start with five carbon you go through
the Pento sugar
transporter transport them inside the
zylos and from there it convert them if
you look at all of them end up in pyu
and then pyu um involve converting them
to acetal and ethanol so this is um you
know just put them all the reaction
together so you can do six carbon and
five carbon depending on the East TR and
also now we use the
modern Gene technology to put one
organism can do both five carbon six
carbon
metab um in a bioprocessing standpoint
you get your raw material then go
through milling and then a big
hydrolysis process you add enzyme amas
to convert the all
the starch into sugar and then you put a
big fermentation tank and use ye to
experiment and then you do product
separation and then you use a co-product
and use dry distill grind for various
purpose like you know feet for animals
and all that so this is just a
bioprocessing steps involved in
converting grin um con con ethanol in us
so just more steps to show you um making
grin ethanol in dry Milling so you need
to grind it up make a liquefaction Stu
to mix with water and and heat them and
make a mash out of that to starch and
then you have scarification step to add
amiz to convert them to sugar and then
you add yeast and to start your
fermentation and then then yeast can do
this job produce ethanol and C2 and then
you do the processing in the other end
where you distillate separate ethanol
and then dehydrate and denature make
sure it is unfit for human consumption
and then you do co-products to use that
like grind for livestock feed and CO2
can be compressed and used in many
different way so this is different steps
involved in gr
ethanol H this is a very mature
technology in the US we have more than
200 Commercial plant if you look at all
these um um plant located most of them
are in Midwestern State this is where
the cor production is and um we produce
7 to eight billion gallon of ethanol
every year from this corn ethanol alone
so this technology is mature and and
it's really doing well
okay but cellulos is a different uh you
know animal itself so there's a lot of
steps involved because of pre-treatment
um of course it starts with the plant
material it's made from plants are made
of cellulose cellulose provides
structure to the plant so you need to
get the cellulose out of this ligin
cellulose complex and we can use any um
plant
biomass agriculture residue um are you
know saw dust paper and paper pulp and
and some crops that is call Energy crop
like switch grass and all that so you
can use any biomass for this cellulosic
ethanol
production the concept is simple so you
get the residue you need to do
pre-treatment steps to delignify remove
the ligin and release the cellulose and
hemicellulose and then you do enzimatic
hydrolysis get your sugar from cellulose
to get glucose from hemicellulose to get
xylos sugar and then the step once you
get your sugar the steps are the same
fermentation and processing and you get
it so that's a simple concept of lnos so
the steps here are this pre-treatment
step and enzimatic hydrolysis step that
is more involved
and costs more and it's one of those you
know limiting steps in this process um
so when you get all these feed stock you
can either do biochemical conversion
which is what I just um showed you and
also you can do thermochemical
conversion so thermochemical conversion
is involve pyrolysis gasification like
Fisher Trove um and then you you get
variety of product through refining not
only ethanol you can buy get butanol oin
gasoline Diesel and then you can use the
the lignin as a Coen for generating
electricity so not only biochemical you
can also use thermochemical conversion
from Fe various feet
stock so it's a the ethanol is good
mainly because it contains um oxygen
adding oxygen to fuel is you know give
you complete fuel combustion and so as a
result um we have 10% of gas um ethanol
is added as a blend in the US currently
Brazil has all the way to E85 85%
ethanol there the advantage of adding
ethanol with the oxygenated fuel is um
you reduce your carbon monoxide emission
and Par matter emission like 50% and
volatile organic compound emission
reduced by 7% so it's like a air
pollution control so it's another
advantage of using ethanol as your
bii so if you look at the projection
from us from cellulosic Material how
much ethanol can be made just from
agriculture waste alone they're
projecting to make 50 billion gallons of
ethanol but potentially you can make 80
billion gallon from all the residues
that they make from various crops in the
US if you use energy crop like switch
grass and other energy crop in marginal
land you cultivate those energy crop and
use it for cellulosic ethanol you can
make 165 billion gallon ethanol per year
okay and that's more than enough for the
country's need and so this is what they
trying to go through in this route and
we had a hiccup in Trump Administration
everything came to a stop but hopefully
this research will go back again and try
to achieve these
goals so the motivation of Bio energy is
three default um you can use the damage
part of the stock and also the residue
that is coming from agriculture
operation for the energy production and
as I said it's less air pollution from
vehicle because of the oxygenated Fuel
and also less burning of crop after
Farmers um harvest the um crop they they
burn the crop for the next year U
plowing next year agriculture operation
so as a result every year
um the fields are burned and you get
pollution air pollution so the you have
motivation to uh reduce air pollution um
and also you have um know energy
security as
well so this just to show you an example
in India every year we have the smog uh
from burning rice crop and also in China
we they have a smog in Beijing so just
to show you pictures of air pollution
from agriculture burning operation this
is the recurring theme every year we
have this problem and nobody's
addressing
it um so the the process involved is not
only just to make ethanol then you had
to do distribution and end use so it's
like a lot of people are involved in
this um steps not only scientists but
engineers and Economist and you know
business people involved so if you're
going to make a more
moreos if you're going to um use land to
produce more fuel crop then initially
you're going to generate more greenhouse
gas because you're going to plow the
land put the biomass in there plant in
there you're going to have some PM
emission going to go up initially but
eventually it's going to you know going
to be neutralize because your land is
going to be continuously used so if
you're preparing the land
initially going to you know have some
pollution
problem so if you look at the biomass
itself it's it's a new crude um sugar is
a crude biomass also is a crude um so it
basically petroleum was one time is a
biomass right all the petroleum products
are started with biomass one time so
same ingredient that made oil is what we
are using now um so the we going to
speed up the process the waste materials
to create renewable energy so you could
start with lot of different crops waste
material biochemical conversion to get
your sugar go through fermentation
catalysis and then you can produce lot
of things not only energy but also other
sugar platform you can put other
chemicals Fine Chemicals you can make
okay so it's a it's a sugar is a new
crude according to American petroleum
Institute so the energy demand is keep
going up worldwide and if you look at it
um um the the the two most country that
use energy high is China and us and then
we have Europe and India are in top four
countries the demand going to keep going
up every year so we need to find new
energy for the
masses so the projection of the energy
is if you look compared 2011 to 2040 the
petroleum uh energy source going to go
down um com from 37% to
31% um and then you're going to have the
biomass going to go up uh according to
the International Energy Research
Institute and so you can just to show
you um you're going to have new
non-conventional sources um is the major
source um in in the future okay because
you're going to deplete your petroleum
Reserve um eventually going to run out
of
it so the imbalance is supply and demand
uh a fuel um so you need to kind of
balance this out and why because people
are moving to big cities and um so this
is how it was in the last century um and
in
1900 now you know cities look like this
so increase in urbanization change in
lifestyle so your energy demand is going
to be high so you need to have liquid
fuel for your transportation currently
that's hopefully that will change
because of electric
car um so solution for all this is
currently it's biofuel okay and probably
50 years from now it could be different
so biofuel is the energy Demand right
now computer is acting
up let's get
this so um let's look at how you can
what are the different biofuel you make
uh you can make a liquid biofuel you can
make gases biofuel not only ethanol you
can make biobutanol um biodiesel
biomethane biohydrogen in the gaseous
form in gas and methane and hydrogen we
call biohythane so these are various
possibilities so currently we're going
to talk about ethanol but we are people
also look into other energy and S
production so one more um uh slide about
advantage of banol is is threefold as I
said is better for the environment
better use of product from Agriculture
and food security and you have um you
know all this carbon neutral better
biodegradation less air pollution
greenhouse gas emission reduce
dependence on oil so you have energy
security and diversification agriculture
so you have taken care of the rural
economy so people in the rural um side
so my computer is acting after um rural
areas can also be taken care of by
participating in this field and you can
have this um energy a lot of different
product and you can also make Fine
Chemicals once you get the sugar you can
you know through bioprocessing you can
make lot of product whatever in
demand sorry about that I think I'm
going to go with this here
um so uh so we already know what is um
this classification of the fuel the
first gen is corn ethanol mainly um and
then we have second gen which is lios
cellulosic third gen biofuel is all
bopes and then fourth one is biohydrogen
okay um if you look at this various crop
that we call that second generation
substrate and you can grow them in um uh
on on and uncultivable land so the the
land that are not usually used for
agriculture you can use them this
biomass crop because they don't need um
better conditions okay you need less
amount of water to grow um it's not
going to interfere with the food chain
so there is no food versus fuel debate
here um there lot of L biomass can be
cultivated in this way so you have more
um resources available okay
um so if you look at the biomass the
problem is this uh getting the ligin
removed and get the these two sugar
containing complex out cellulose and
hemicellulose okay so if you look at the
biomass itself the major component of it
is um uh ligin hemicellulose and
cellulose and depending on the crop and
the biomass it varies so yeah up to 50%
in some cross crop cellulose so these
are the three major ones so we can use
the sugar and then we could use liin for
other purposes so in my case I'm going
to use ligin to make microbial lipids
okay so how how this is all started in
big way it started because in during
Bush Administration second Bush um we
had this oil price went up almost $110 a
barrel so the President Bush promoted um
cellulosic ethanol concept and then and
uh President Obama took it forward and
gave a lot of incentive lot of companies
put commercial plan and then politics
change and now everything is not doing
well uh so hopefully it will do well in
the future so I want to just give some
basic facts about this um so we do not
need energy but we need services that
energy provides uh those services are
heat light mobility and Mobility you
need this liquid field currently right
um so energy has fundamentally different
quality so all BTUs are not created
equal
um our society will literally stop if
there is no liquid field currently so we
need there is a demand for liquid field
so liquid Fields
um not energy are required for Mobility
for the for at least few decades at
least you know electric cars are picking
up but at least for you know few more
decades we need liquid f so just to show
you the whole society will come to stop
if if you don't have liquid Fiel
available um if you look at all energy
carriers um um have not have say um do
not have equal strategic import because
of if you look at different energy like
coal us and China have plenty of them
right and domestic resource and natural
go gas you have a lot from Canada and
Mexico and petroleum um during this
energy crisis 60% was imported but Curr
currently us is um energy exporting
country because of um you know drill B
we drill um last few years we've been
drilling everywhere even in National
Park and um and also fracking uh the new
way to get Shale um petroleum from sh
so we are producing more but but people
are predicting this is not going to last
long okay um so it's going to that's why
we need to have a alternative because
petroleum can undermine a lot of thing
climate security Economic Security and
International Security and World
stability so so all countries need to
have their own energy um Independence so
the biofuel and liquid um cellulosic
ethanol is a way to go um according to
many many many scientists
okay um so the Cal is the you know
alternative to
petroleum for these reason as I said um
National Security greenhouse gas
reduction and economic advantage and
also uh the the presentation of
emphasize in this particular talk I'm
going to talk about cellulosic ethanol
from various biomasses
so again go through one more time the
schematic this is the schematic for
ethanol from corn you get corn kernels
and um go through the um hydrolysis and
um get sugar and fermentation process
and and they do distillation drying and
ethanol get ethanol out and then you get
co-product that goes to mainly animal
feed in us and this is sugar cane like
in Brazil H sugar comes out of sugar
cane and goes straight through this prod
almost the same you have this because of
the sugar you don't have this hydrolysis
step for the sugar
can
um and then if you look at the um um the
cellulosic ethanol have a lot of steps
involved pre-treatment hydrolysis and
then once you get the sugar the process
is the same there is no difference okay
and then um you can also include the
thermochemical conversion so you get the
after pre-treatment you get the solids
and use this thermochemical conversion
to get heat power and and Fuel and
chemicals
okay
um if you look at all commodity um there
are the price of this energy based on
raw material how much it's going to
cause the feet stock and the processing
right so when you started in gas in
initially the cost was getting the raw
material and um and then the processing
cost keep going down in the future so so
the same thing we are predicting for
lnos cellos lnos ethanol production um
how much is going to cost the raw
material and what is the processing cost
the processing cost going to uh going to
go down because of Technology because of
research okay um so we have this Mar to
play with depending this um cic ethanol
is tied to the gasoline price um so the
gasoline price fluctuate if you look at
last 20 years you know so we have this
big fluctuation so we can uh so we need
to have a stable price um that's why
these companies that are set up to make
this are failing currently because
petrol is cheap currently it's really
cheap
here so he how the price for the raw
material and then the processing cost if
you look at the um initial uh petroleum
energy um started uh there was the cost
for the oil was less but processing was
high and then all this refining uh
research reduced the cost of processing
and then because then the dwindling
energy Supply so now your raw material
is going to be h going to cost more and
your processing cost less so so this is
the same scenario we are looking at in
cellulos initially processing going to
cost once the technology mature your
processing cost going to go down but the
same way it played for petroleum
industry so just to show that that's
what happened in Brazil in Brazil when
it started this whole um ethanol
production there energy independent
country the ethanol cost went down um
because of the processing um cost went
down um because of research and
Technology um to make this
possible so we are we are advocating the
same principle
for Theos cellulosic ethanol production
as well so here is today's cost you if
you look at it the processing is more
because of enzy cost pre-treatment steps
all that going to cost more um biomass
prices less but in the future we are
predicting because of technology and
science this cost going to go down and
and the biomass is going to be the cost
going to go up production of the biomass
going to go up okay so very similar
principle is what is expected in in the
future so when when the cic ethanol was
at Peak here like 10 years ago we had
these commercial companies set up shop
in us they were producing commercially
from um corn store and um um they used
different pre-treatment the one of the
successful one was aex um pre-treatment
I'm going to talk about in a minute uh
but currently all of them are bankrupt
because of political change and oil
price so so so we had so many success
story and then
um again it it follows the price of oil
so so if you look at the the process
itself uh you you get your biomass and
you have the harvesting storage size
reduction and then you have the
pre-treatment step and and you enzyme
you need to have enzyme production and
then Hy hydrolysis step and you get the
sugar and then you get a fermentation of
sugar to make your ethanol and then you
have the residue and the waste treatment
so this is the complete picture of how
all this commercial company um you know
did this process okay but the
pre-treatment Step was different for
different companies
okay so this is the whole view we get
the harvesting storage and so um size
reduction of biomass transportation to
the field and that's that's an all cost
involved then pre-treatment and um then
the process these steps are um almost
similar but the pre-treatment and entic
stuff are different depending on the
different company so one of the
successful pre-treatment versus FX which
is the ammonia fiber expansion method uh
so we use they use ammonia and for tree
treatment sta in a reactor so when you
heat the biomass at 100° with
concentrated ammonia you have a um rapid
pressure that release the end treatment
as a result you get the sugar release
but there is no other Inhibitors that's
one of the advantage of ax there is no
fural for five hydroxy methy for no
Inhibitors produced during FX process so
this is very successfully used
commercially and people are um hoping
this is the way to go in most of the
Midwestern states in the US so they did
the lot of comparison there's a price
for using dilute acid pre-treatment h
hot water and then here's a effect a141
per gallon of
ethanol and then you can compare with
other um tree treatment process so this
this method um is commercially used in
three different companies and very
successfully and the cost also went down
because you can reduce ammonia and
recycle ammonia then you can also reduce
Capital cost um doing affx analysis I'm
going to do a couple of modeling and and
see how this price is going to go down
okay currently they're pricing
a141 per gallon of ethanol during FX
process okay and this is the enal model
National renewable energy lab um and
then if you look at the price in the
future by Consolidated bioprocessing you
can make ethanol 62 cents per gallon
this is a model they're predicting and
also go below 60 cents you when the
technology mature okay so they're hoping
this model will compete with the oil
petroleum price and this is going to
take off in the new Administration um
coming
up um if you look at what is happening
because of the inexpensive ethanol you
have um other problems we have um
environmental Improvement possible um
rural economy is in US
all the rural people are really dying
out though there's nothing to live for
so they're hoping this technology will
revive the economy in the rural area and
um also you have more food for the
animal because of the distill and the
grin that comes out through the process
you get animal feed possible so these
are additional um uh incentive to
develop this technology and people are
pushing for the is in US
currently um so the Michigan State
University come up with this plan um if
you if if this
technology has to be you know taken up
Nationwide and also internationally you
need to have this set up in small rural
areas like like a Cooperative so you can
reduce the transportation cost so to get
the biomass to the plant so you set up
this Regional biomass processing center
and then get your pre-treatment going uh
and then you can make um a lot of
product you get your high value um users
and you can get um you know the waste
whatever waste you can do Coen per power
plant and then you have a bio refinary
to make your ethanol so this this is
available in in the Michigan State
University website and this model is
what they are predicting is the future
for this industry
then there are certain myth out there um
I just want to clear some of those uh so
myth number one is ethanol has negative
net energy um in reality gasoline's net
energy is worse than ethanol if you
compare this metric is not is not
relevant okay another myth is ethanol
will drive up food prices because the
debate of food versus f is a little
complicated there's no easy sound B for
this um but cellulosic ethanol will you
know will not compete with food because
we looking at the biomass and waste from
agriculture operation and it reduces the
greenhouse gas and air pollution and the
another myth is ethanol is bad for the
environment what we are comparing to
what you're saying ethanol is bad uh if
you look at K ethanol it's actually SEC
gasoline um for most metrics cellos EOL
is will be better because again we using
base product mostly ethanol will always
cost more than gasoline currently it is
true but ethanol from corn we are making
um in the commercial sector they're
making a120 a gallon uh from cellulos as
as a model indicate you can make 60
cents a gallon so so these are what they
are hoping could happen to this industry
and going to pick up again okay so the
another part was um getting this ligning
out of the um out of the biomass to get
the sugar so mostly people are working
on pre-treatment of chemicals like acid
alkaline ionic liquids and um but we can
also use
biological processes using fungi to
remove the ligant and so you can combine
these two methods uses little bit of
acid treatment also combined with the
fungal enzyme to remove the ligning so
so if you look at it there's a lot of
fungal enzyme lactases and peroxidases
could be used and um so just to show um
why L is there in the first place to
protect the plant and now we can use
this um fungal enzyme to you know use
this in this
technology so we can uh use this um
pre-treatment and you can use this uh
fungal as a biological pre-treatment
method and try to reduce the price of
the whole technology so I'm going to
talk about that in my
research uh just to show you there are a
lot of pre-treatment out there and these
are the acid alkaline and then physical
chemical method and enzimatic Method and
the two commonly used method the one is
the ammonia fiberx
which is already commercialized in us
and ionic liquid pre treatment is also
good but it is expensive currently but
commercially not um possible compared to
FX
process and I am advocating biological
pre-treatment using um various fungus
because this fungi produce a lot of
these enzyme manganese peroxidase liin
peroxidase and La cases um so we can use
uh some of these fungi that are high
efficient um biologically they can
remove liin
okay
um it's also um you know it's very
targeted these enzyme only attack line
so the sugar is not degraded uh so if
you stop the reaction then you can get
the whole sugar and you don't have any
Inhibitors that is produced during this
process uh you can easily recover the
enzyme and reuse the enzyme so the cost
will go up another advantage using
biological um pre-treatment using enzyme
is this water uh um reduction so if you
look at all this method um you're using
a lot of water in pre-treatment but if
you if you use the biological method the
water usage is very very less all right
so you produce less waste you consume
less uh water in the whole processes so
this is the way that I advocate in my
research to use biological pre-treatment
using fungal
enzyme um just to show you the enzymes
involved a lot of different enzymes you
can use in in your in the process but
generally there are all Lac cases
manganese
peroxidase all those enzymes are
commonly used
okay and we can also people are looking
in now enzyme from different sources
like termite um different crbs insects
and they're looking at microbes inside
their guts because these are all biomass
processing insect they eat these plant
material and so they're trying to get
some novel enzymes So currently the
triodes is the the one that industrially
used for enzyme for the getting
hydrolysis but also looking at different
um sources to get novel
enzy so again uh you need to get the
ligning out release this um cellulose
using the cellulose enzyme so you can
get them from various sources and when
you do that and this is how the
biological uh pre-treatment look like uh
you get your
biomass uh you get your um process your
Bas in the powder form or whatever form
that is CA effective and then use this
enzyme to
delignify you can use the whole fungus
or you can use the enzyme out of the
fungus and process it and then you that
will get your sugar out and then you
have this bio ethanol production from
various sugar and you get your ethanol
from lios cellulosic so uh my research
is to combine one of those chemical
method and the biological method to make
it more efficient so that's what I'm
going to talk about from my personal
research so just to show you if you use
these enzyme you can see the picture raw
biomass and pre-treated biomass you can
see what these enzyme can do to get the
ligning out and make the structure uh
after it is pre-treated with this
enzymes so um less water use it's a
better method and can be recycled is
also cost effective
okay so with this broad overview
on uh how this cor ethanol solic ethanol
and some commercial plan already in the
US they started and then now bankrupt
and hopefully they'll go back because of
a um the bioprocessing cost is coming
down through research okay so I was
involved uh seven eight years ago um uh
in this uh field I my research was
funded by Department of energy um so I'm
going to share some of my uh research
with you in this field okay so as I said
my research was to reduce the cost so
you want to combine the biological
method with the chemical method for
pre-treatment so you can reduce the cost
and make it more economically viable
this process so we I don't have to go
through this we already talk about why
we talk we use the fuel liquid fuel
because it's mainly for transportation
purpose and then this liquid fuel um
from the energy source we uh it's also
carbon neutral once this process mature
once you get new land into cultivable
land um then you can see most of them
will be carbon will be recycle and
become carbon neutral and release less
CO2 and less greenhouse gases okay
um so this is currently we account for
1% of world energy from demand from
biofield I'm hoping this this number is
going to go up when when new government
takes up uh in US uh so we need to
make
moreos ethanol more economically viable
and feasible okay so this was a old
figure when I started the research the
doe wanted to have this 60 billion
gallon ethanol production and they
wanted to get this
$233 at that time the oil price was high
H but now you know it's now they want to
have a dollar2 so so that's a new
figure so I was involved in um the sugar
cane uh as a raw material because
Louisiana is a sugar cane producing
state one of the major crop is sugar Su
cane and um so we have a lot of this
after sugarcane Harvest we have this
Leaf litter on the ground um currently
they burn it so you can collect this
leaf and use it for L ethanol production
we can also use energy cane so there are
some sugar cane that are classified as
energy cane because it has more um
cellulose and
hemicellulose and this can be grown year
around and it can be grown in um non
aable land because they they disas
taller and there a lot of advantage of
using this energy
cane so just to show you there are
different type of sugar cane this is
commercial sugar cane this energy can
one energy can two you can see the
difference in cellulose semic cellulose
and sugar content in this crop okay so
there are varieties um different
varieties we used I'm going to talk
about the one variet
um that for my research because of
time um so we we collected this type S
which is energy can two from the USDA
research station um for our research we
brought the lab and dried it and process
it um to do your pre-treatment Stu again
just to show you um we need to remove
the ligning get the cellulose and
hemicellulose out you can use chemical
or biological free treatment
so once you get cellulose it's 100% um
glucose right so once you get the hyd
hydratic steps the cellulose can be
broken down to get your
sugar um and then when you remove
hemicellulose you have this pentos and
hexos that makes hemicellulose We
combinely call xylos we get all the
sugar released that could be used in
your fermentation process and then we
have uh this lignin all right uh ligin
could be in the other process they use
in Coen uh but I am proposing to use
this liin using specific bacteria to
make
microbial lipids which could be used as
biodiesel so uh the my research is using
everything uh from the biomass sugar
both pentos and EXO sugar and also um
the ligant for microbial lipid
production
uh this is just a cartoon to show how
the steps work uh get your pre-treatment
get your entic digestion fermentation
distillation um so what type of enzyme
to use to get your um sugar released
okay so our objective is to try this
particular type to energy can when I
compare various pre-treatment combined
with biologic iCal treatment optimize
the condition and then um how best we
can use the biological and one of these
pre-treatment method in our
research um so we started in small scale
um and we used the acidfree treatment um
and using sulfuric acid weak sulfuric
acid and then we you know processed it
and then we get the Lig out of the um
pre-treatment steps then we use en start
to get the the glucose out for
fermentation and then we use various
fungus we use the whole fungus white rat
fungus brown rat fungus and then use
this Sol State fungal treatment and this
method was advocated by Dr ranish suari
when she visited here as a full bright
scholar so this is from her research uh
so we started using this method uh whole
fungal inoculation and look at softening
of the
Li and then they have entic
scarification step we use this different
enzyme for cellulosic enzyme and xylinas
for hemos so we get both um pentos and
hexos out of this biomass
okay uh we used the first East and then
we used the recom and eoli this recom e
like can ferment both pentos and and
hexic sugar so we can have better eeld
okay so just to show you uh just a
pre-treatment alone comparing um
different acid pre treatment so if you
look at you know 4% versus 1% um
4% of acid volume by weight work better
um in terms of ethanol we we not using
anything else no entic stuff just
pre-treatment whatever sugar that come
out um so you can see that um between
three and four there is no significant
difference in ethanol eel so we
advocated 3% sulfuric acid could be the
better way to go for this pre-treatment
stuff for this particular biomass and we
also want to compare whether alkaline
will work uh so alkaline pre-treatment
we increase the pH uh from you know
eight all the way to 13 and you can look
at 13 work better but then there's no
difference between 12 and 13
statistically so we stick with
PH2 this is biological so you can use um
uh individually the white rod and brown
rod puni in in a solid state
fermentation or you can combine these
two together so when you put these two
fungi together your ethanol yield was
better so so we advocated putting this
fungi together to soften up the
ligin so just to show just without doing
anything how much sugar is released in
this process you can see glucose and
xylos yielded various pre-treatment and
you can see the 3% 4% acid there's not
much different and and then the fungal
pre-treatment when you combine these two
fungi you have better sugar yield in our
lab experiment okay and then we we went
with
a step of how we can combine these two
biological and um acid free treatment um
so we the idea here is when you collect
biomass in a large scale you have to
store the biomass anyway right so during
the storage process you can use this two
fungi to spray it on the uh bi Mass uh
uh and then after you know couple of
days then you do the pre- treatment step
so in this way you can use less acid um
so this is just to show you when you use
the ethanol your acid pre-treatment uh
your concentration is less you don't
have to use 3% you can use 1% acid so
your acid cost going to go down so that
that's experiment we did just to show
you you just use this two fungi together
for two days um and then you put those
fungi in acid treatment and come compare
various acid treatment so without fungi
you need 3% acid with fungi two day
storage you need only one person acid
okay so this makes sense so this will be
economically viable process
okay um so that yield was not that good
when he did did the ethanol so then we
did the genetic
engineering uh we took this eoli and and
then we just did some knockout
experiment we put some
zamonas gene into this
eoli and try to get better yield of
ethanol so want to get um theoretical
yield of ethanol so those thing are
without genetically modified organism
just the East now we are using this
recombinant eoli just to show you so we
did some genetic knockout so we knocked
out some of the genes in this eoli um
because this eoli can produce from sugar
variety of product not only ethanol it
also produce acid succinate lactate so
we knocked out the genes that produce
all these product by only allow the gene
that produce ethanol so in this way when
we have this come in an eoli you can
make get 95% yield okay so we got this
eoli to work um and and after we did
this combined pre-treatment with the
fungus and the acid and just to show you
the the process uh so here is your xyo
sugar going down here is your growth of
your eoli and organic acid when there is
no knockout when you knock out those
genes you don't have any acids you
produce everything one product ethanol
so this is uh you know almost
theoretically getting from this
process H so with with that we scaled up
this process and we used
Bas which is coming out of the solid
waste coming out the Sugar Mill um to to
do the same step combined with the fungi
uh two days of storage and then you use
one person acid and then use this recom
Coline okay and we used
um variety of enzyme to get the sugar
out of this because the Sugar Mill they
didn't care they wanted to you know find
out how much maximum sugar they can get
so if you look at it we use lot of
different enzyme we can live with couple
of enzyme but we wanted to show uh
complete use of of the biomass so so in
this case we use lot of enzyme but I
will Advocate we can live with maybe
couple of enzymes so we don't have to
worry about you know arabos those are
all very small amount in the in the
biomass we want to use all the sugar
that comes out of hemus so we scaled up
to 5 lit and then to 100 liter um I'm
going to show you some results from that
experiment using recombinate so here is
the the biggest scale experiment and you
can see um after the processing of the
bagas with two days of um fungal
treatment 1% of acid um your glucose
yield and xylos yield uh in their
fermentor and they all went down because
the EOL can use both sugar and both
sugar went down and then eolic growth
went up they're using sugar to grow and
then the total sugar when combine these
two sugar the total sugar go down and
then your ethanol yield coming up okay
so in this 100 lit experiment we did not
get theoretical yield yet we still have
to play with this process a little bit
uh we we got some hiccup so if you look
at it we didn't get
4.49 mole we only got half of it so we
need to improve this process further so
this particular experiment um we combine
two pre-treatment so the the the logic
behind is when the companies Harvest
this biomass they have to store it
anyway uh so during the storage they can
use this two fungi and then they can use
less pre-treatment so that will save
some money so that's the concept behind
this research okay so we used the pentos
and um exos sugar now we have Li right
so we wanted to use Li so the idea here
is to find an organisms that could use
all polyphenol that come of ligin come
out of ligin and then you know convert
them to microbial lipids and could use
as biodiesel right so so we can use
every part of the
biomass so um just to show you why we
went on this microb lipid angle again
the same story we have we got a have the
energy we need the biomass uh is one of
those way to go I don't want to repeat
this again um this is again how much
energy can be produced from biomass and
this is a US goal in 2022 and they're
not going to achieve it um currently
we're producing 10 billion because of
the government change the Trump
Administration completely shut down this
so we are not making this so hopefully
this yield will go up in the future the
US um so you can the process is to go
through fermentation and make biofuel
and then in the
other way how they make bio diesel is
transesterification from plant material
right so we want to make microbial
liquids as a so way to go make
biodiesel um so just to show you the
government change screwed up the whole
um process of Bio energy in the US so
the last four years the energy policies
changed and drill baby drill regulations
are relaxed for petroleum industry and
completely shut down
liic activity in the US many commercial
ventes went
bankrupt so these are the company that I
pointed out before um all most of them
uh went bankrupt this one is still
operational duon in Iowa is still
operational uh the fulcrum is still
functioning and the other companies went
bankrupt so because of government change
in
policy so again uh the biodiesel concept
is simple so you get your um uh biomass
and you do your pre- treatment uh and
then you you go through this
transesterification process and you get
your biodiesel and then you get glycer
in as your byproduct and um so the steps
involve trans recation steps involve
your methanol and your glycerol is one
of the product you get variety of
different uh lipids that come out okay
as a biodel so the organism we chose was
this oogenus micro rocus rocr um rocr
opas is also one of the good one so we
chose Roc rocoss which is because it's
also bio remediate
microbes it is very versatile so the
this organisms can convert um the sugar
into um final compound into um lipid
inclusion bodies and you can easily
separate this um so it doesn't involve
any big distillation to get him out so
it just open the cell you get the lipids
out the easy bioprocessing of getting
biodiesel
uh so we uh we started out small and and
and did a pilot plan study on this in
Mississippi State University with the
chemical engineering department there um
I'm going to show you how we started by
using the ROC caucus with um first
glucose and then we use the
ligin polyphenolic compound okay um
again the concept of using is um this
way if you go through this uh bio
microbial Liquid Concept This is highly
efficient compared to growing plants and
getting the
biod and you can you know grow
microorganisms really easily in a big
tank and and you make more out of this
compared to growing
plants
so okay sorry about that and my computer
is acting up today I'm trying to get
this again started
[Music]
so um just to show that this is the
concept um you get the the L paste get
I'm sorry Rich the screen is not yet sh
oh it's not showing yes okay I think I
have a problem with there today do you
want us to show your
presentation yeah you have it with you
yes uh Gund can you help
us yes
okay yeah start with this um slide that
shows the rocus
[Music]
I have
[Music]
it so you canect yeah I can see
[Music]
here yeah go down all the way to the
micro liquid part
in the slide 110 or something G yeah
something like that
[Music]
yeah and while waiting uh you can also
ask the question in Indonesian yeah we
will translate it to English for
Professor R oh yeah yeah that's good
sorry about this technique glitch it's
okay it happens all the
time it work last three classes it
worked I don't know what happened
today yeah this is good so this this
slide I was just mentioning about the
comparing growing plants and then
getting your
SNR using the microbial route is nine
times more efficient okay just to show
you you know if know when you want to
grow plants and you're going to have
this inputs and all that but you don't
have that in using microbes okay okay
next
one uh so the concept is simple you get
the ligning when you do your biomass and
the liant is mostly phenol and this
organism can use phenol as carbon source
to grow and then and and when you um
grow this and they're going to you know
with high amount of carbon in during
when you have high carbon it's going to
convert them into this microbial lipid
and then the lipids can be easily
removed and you get biodiesel okay next
slide uh so we started out just to show
the concept with the glucose so we
didn't use the ligant okay so this this
produce the pigment and just to show how
much biodiesel it can make just using
sugar as a raw material okay next next
one
um so we did the analysis using um you
know lipid extraction BL and dryer
method and freeze dry pallet to look for
biomass how much biomass is produced
okay next
one next
slide uh if you look at it here you can
see the lipid
production and the glucose consumption
um using different concentration of
glucose 10 gram per liter 20 gram per
liter 40 gram per liter so the more
sugar we had the more liquid production
we had in our system so just to show
this concept work we have the liquid
produced from sugar so now we want to
substitute the sugar with the phenol
from Lin okay next next next slide and
just to show these organisms once we
grow them on sugar and we got this
um the oil droplets and and we can
process them this micro lipid can easily
broken down and can be released so this
is our organisms in
prot okay next
one next
slide can you forward next
slide okay so what