Sugar Cane industry

drawerbeamerBiotechnology

Dec 6, 2012 (4 years and 8 months ago)

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1

Utilization of Waste in

Industrial (White) Biotechnology

White Biotechnology

2


is an emerging field within modern biotechnology that
serves industry.



It uses living cells like moulds, yeasts or bacteria, as well
as enzymes to produce goods and services. Living cells
can be used as they are or improved to work as "cell
factories" to produce enzymes for industry.



White Biotech can help realize substantial gains for both
environment, consumers and industry.

Biotechnology

Green

Agro
-
Food

Red

Health Care

White

Industrial

Health

Unmet Needs

Economy

Sustainability

Using nature’s toolset

4

Industrial (White) Biotechnology

Sugars

Biofuels

Biomaterials

Biochemicals


Cell factories

5

The IB Value Chain

Biofuels

H2

Ethanol

Sugars

Feedstocks

-
Renewable

-

Fossil

Biochemicals

Food Ingredients

Pharmaceuticals

Fine Chemicals

Biomaterials

Polylactic acid

1,3 propane diol

PHAs

Bioprocesses

Bulk

Fine


Cleaner and more (cost) efficient ways of making:



Faded jeans


Detergents


Plastics


Vitamins


Antibiotics


Fuel


Biosteel


Biobatteries


DNA computers

Industrial Biotechnology


Reduced environmental
foot
-
print up to 20


60 %




Added Value of

11
-
22 billion


per Year


IB: three
P’s

go hand in hand


sustainability

Profit

People

Planet

8

The IB Value Chain

Biofuels

H2

Ethanol

Sugars

Feedstocks

-

Renewable

-

Fossil

Biochemicals

Food Ingredients

Pharmaceuticals

Fine Chemicals

Biomaterials

Polylactic acid

1,3 propane diol

PHAs

Bioprocesses

Bulk

Fine

Strong points Europe



Enzymes



Biochemicals

9

The IB Value Chain

Biofuels

H2

Ethanol

Sugars

Feedstocks

-
Renewable

-

Fossil

Biochemicals

Food Ingredients

Pharmaceuticals

Fine Chemicals

Biomaterials

Polylactic acid

1,3 propane diol

PHAs

Bioprocesses

Bulk

Fine

Biomass

Bioenergy

B & B

10

Developing a Strategic Research Agenda and Roadmap (1)

Main R&D objectives



Strain, biocatalyst & process optimization



Novel and/or improved functionalities and products

11

Developing a Strategic Research Agenda and Roadmap (2)

Research & Technology areas in IB



Novel enzymes and microorganisms


metagenomics



Microbial genomics and bioinformatics



Metabolic engineering and modeling



Performance proteins and nanocomposite materials



Biocatalyst function and optimization



Biocatalytic process design



Innovative fermentation science



Innovative down
-
stream processing



Integrated biorefineries

12

Novel biotechnological processes

for production of polymers,
chemicals, and biofuels from waste


Background

13

13


Ecological

reasons

to

promote


White

Biotechnology

:


Global

Warming,

Green

house

effect




Rio

Declaration

on

Environment

and

Development


June

1992
:

Broad

consensus

to

switch

to

alternative,

sustainable

Technologies





Principle

4
:



„In

order

to

achieve

sustainable

development,

environmental

protection

shall

constitute

an

integral

part

of

the

development

process

and

cannot

be

considered

in

isolation

from

it
.





Rising

Prices

for

mineral

oil
:

Economic

necessity

to

promote

technologies

independent

from

the

availability

of

fossil

feedstocks



Major

Drawback

for


White

Biotechnologie

:

Costs

for

Raw

Materials



Solution

Strategy
:

Utilization

of

Waste

Materials

for

Production

of

Biopolymers,

Biochemicals

and

Biofuels


14

14

140

130

14


June 2008: Price surmounted 130 US
-
$ per barrel


July 2008: Price surmounted 140 US
-
$ per barrel

Need for

White Biotechnology


for Production of
Biopolymers, Biofuels and Biochemicals?

15


Target Areas

: Final Products from Conversion
of Waste Materials via

White Biotechnology


Major substrates for production of
biopolymers, biofuels and biochemicals
:

Monosaccharides:

Glucose,
Galactose, Fructose, Xylose,
Arabinose

Disaccharides:

Sucrose,
Lactose, Maltose, Cellobiose

Polysaccharides:

Starch,
Cellulose, Lignocellulose

Organic acids

Lipids

Alkohols:

Glycerol, Methanol

Industrial producers

of Waste streams:

Proteinaceous
materials (Peptides)

Biodiesel production
: raw
glycerol phase, low
-
quality biodiesel
fractions

Dairy Industry
: Whey

Wood processing industry,

Paper Industry

Additional agricultural
branches

(e.g. straw from rice,
mais etc., olive oil production, palm
oil industry, sugar beet industry)

Slaughterhouses and Rendering
Industry
: Meat
-

and Bone Meal, slaughter
wastes

Biochemicals

(Fine chemicals,
Organic acids,
Antibiotics, Aromatics,
Surfactants, Solvents,
Chiral Synthons)

Biofuels

(Bioethanol,
Biodiesel)

Biopolymers

(PHA, PLA)

Catalytically active
Biomass

for Production
of Biopolymers, Biofuels
and Biochemicals

Sugar cane industry
: Molasses, Bagasse

Final Products:


16

Brazil: Integration of Biofuel & Biopolymer
Production into Sugar Cane Industry: Actual and
Potential Utilization of the waste streams

Sugar Cane


Milling

Extraction

Raw Juice

Crystallization

Molasses

Saccharose

Steam and
electrical power

Combustion

Bagasse

Hydrolysis

to
Glucose
and
Fructose

Convertible Sugars

(Glucose, Xylose, Arabinose)

PHA
Biopolymer
Production

Fermentative
Conversion to
Bioethanol

1.) Production of
catalytically
active Biomass

2.) Production of
PHA biopolyesters

Hydrolysis

Biofuel
Production

Destillation

Bioethanol

Higher Alcohols

(Butanol, Pentanols)

PHA
Biopolymers

Downstream
Processing:

Extraction of PHA
from biomass


Residual Biomass

Hydrolysis

to
peptides
and
amino acids

Selection of production strain!

Fibers potential
filler
for PHA
-
based materials?!

Extraction
solvents!

180.000 t/a

30.000 t/a

10.000 t/a

561.600 t/a

32,4 GW/h / a

395.000 t steam/ a

52.575 m
3
/a

2,160.000 t/a

17

PHB INDUSTRIAL S/A


S
ao Paulo,
Brazil

View of the PHB Pilot Plant for 50 t/a

Production strain:
Cupriavidus necator

DSM 545 (formerly
Wautersia eutropha
)

Intented industrial scale production of PHA: 10.000 t/a

Production of PHB
homopolyester and Poly
-
3
-
HB
-
co
-
3HV copolyesters
from sugar cane saccharose;
autarkic energy supply!

Basic research: TU Graz,
Austria

18

Whey from Dairy Industry


a versatile Feedstock
for Biotechnology



Application of
Whey lactose

(
D
-
gluco
-
pyranose
-
4
-
ß
-
D
-
galactopyranoside)
from dairy industry: animal feed,
sweets, food processing, baby food, laxatives,
pharmaceutical matrices


But:

annually 13,500.000 t of Surplus Whey in
Europe (620.000 t lactose)!


Ecological problem;

polluting whey partly disposed
in sea


2001: EU


project
WHEYPOL

(G5RD
-
CT
-
2001
-
00591):

application of surplus whey from Italian dairy
industry as substrate for PHA biopolyester production

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From Milk to Whey towards PHA
Biopolyesters



Pasteurization

Transformation

(enzymatic or acidic)

Skimming

Full Fat Whey

Pasterization

Concentration

WHEY CONCENTRATE

Ultrafiltration

Whey
Permeate

Whey
Retentate

Curd cheese


Desalting ?!


(necessity depends on
production strain)

α
-
Lactoglobulin (2 wt.
-
%),
ß
-
Lactoglobuline (9 wt.
-
%); Lactose (15 wt.
-
%)

20


21 wt.
-
%
Lactose

(81% of the entire lactose from milk)

(ca.
620 000 t/a in EU from surplus whey!
)

Skimmed Whey

(ca.
13 500 000 t/a in EU surplus!
)

(ca.
2 700 000 t/a in EU surplus!
)

Storage

Storage

MILK

Lactose Hydrolysis to
Glucose and Galactose ?!


(depends on production strain)

1.) Production of
catalytically
active Biomass

2.) Production of
PHA biopolyesters

Yield PHA/C
-
source = 0,33 g/g:
ca.
200 000 t/a PHA

in EU
from surplus whey possible!!

20

Different Routes from Whey Lactose
to Biopolyesters

(Koller et al., 2007)

Whey Lactose

Hydrolysis

towards Glucose
and Galactose
for Production of
PHA

Direct
Application

of
Lactose (sufficient
ß
-
Galactosidase
activity of
production strain)
for production of
PHA

Bioconversion 1:

via
Lactobacilli
from Lactose to
Lactic Acid

Bioconversion 2:

from Lactate to
PHA

Polylactic acid

(
PLA
)

(www.igb.frauenhofer.de/
WWW/GF/dt/GFDP 21
Molke.dt.html)


Conversion to
Lactic acid esters



Green Solvents

Pyrolysis

Unsaturated compounds

(Crotonic acid, 2
-
Pentenoic acid etc.)

Synthons for chemical synthesis

Lactones

21

Alternative Biotechnological Products
from Whey Lactose


Bioethanol:

Golden Cheese Company, California (19.000 m
³

Bioethanol/year) (For Europe: Surplus whey would yield 290.000 m
³

Bioethanol/year)
(www.ethanolfra.org/pr010201.html)


Antibiotics:

e.g. Bacteriocin
Nisin

(polycyclic peptide antibiotic from
Lactococcus lactis
) against highly pathogenic food
-
spoiling bacteria
Listeria monocytogenes

and
Clostridium botulinum
(Hickmann, Flores, Monte Alegre, 2001)


Sophorolipids:

Emulsifiers and Surfactants for pharmaceutical,
cosmetic and food industry; chemically: sophorose derivates linked to
hydroxy fatty acids


Two

step process: Yeast
Cryptococcus curvatus

cultivated on
whey permeate, accumulates single
-
cell
-
oil (SCO) from whey
lactose. SCO is converted in a second step to sophorolipids by
Candida bombicola
(Daniel
et al
., 1999))

www.lipidlibrary.co.uk/Lipids/rhamno/image006.gif


www.profoodinternational.com


22

The Increasing Amounts of Biodiesel


Legislative Situation by the European Commission:
Shares of Biofuels [%]:


2005: 2%


2010: 5,75%


possibly up to 20% until 2020 (8 * 10
10

Liter/a in Europe)


2005: Production of 1,925.000 t in Europe (
=
192.500 t
glycerol
)


2008: 2,649.000 t in Europe (
=
264.900 t glycerol
)


Austria: 2006 Production of 121.665 t Biodiesel; 2007:
241.381 t (+98%!!!)


23

G
lycerol
L
iquid
P
hase: Waste from the Biofuel
Production for the Production of Biopolymers

WASTE LIPIDS

MeOH (EtOH)

OH
-

Transesterification

Mixture Biodiesel
-
Glycerolphase

Separation

Washing, Dewatering

BIODIESEL (RME)

GLYCEROL LIQUID PHASE
(GLP)

Degreasing

Demethanolization

1.) Production of
catalytically
active Biomass

2.) Production of
PHA biopolyesters

Yield PHA/C
-
source = 0,33 g/g:
ca.
88 000 t/a PHA

in EU
from surplus GLP possible!!

Biotechnological Production of PHA
Biopolyesters

Downstream Processing

PHA
Biopolyesters

Residual Biomass
(Proteins, Lipids)

e.g. Waste Cooking Oils,
waste animal fats

typically 2
-
4 wt.
-
% of biomass

Some lipids: direct
application as feedstock!

Low
-
quality biodiesel fractions:
excellent feedstock for PHA
production!

24

Lignocellulosic Feedstocks



Occurence

of lignocellulosic waste:


wood residues (including sawmill and paper mill discards)


municipal paper waste


agricultural residues (including
corn stover
, rice straw and sugarcane
bagasse
)


special energy crops


Amounts:

non
-
wood lignocellulosic straw alone is estimated with 2,5*10
9

t/a



Composition of Lignocellulose:

Carbohydrates

Lignin (
Methoxyphenylpropane
)

Cellulose fraction

Hemicellulose fraction

Monomer:
Glucose
(Hexose)

Monomers:
Xylose, Arabinose

(Pentoses)

+

25

Biotechnological Utilization of
Lignocellulose


Obstacle:

Lignocellulose has evolved to resist
degradation and to confer
hydrolytic

stability

and
structural robustness to the plant cell walls by
crosslinking between the carbohydrates and the
lignin via ester and ether linkages


Focus of research:
UPSTREAM TECHNOLOGY
:
Enhanced lignocellulose digestion and the
development of
EFFECTIVE ENZYMES

for the
degradation of cellulose and hemicellulose into
glucose and pentoses are the prerequisite for an
efficient production of the desired bio
-
products


26

Composition of Different Lignocellulosic Materials

Cellulose

[wt.
-
%]

Hemicellulose
[wt.
-
%]

Lignin

[wt.
-
%]

Corn cobs

42


45

33


35

10
-

15

Corn stover

35

25
-

38

35

Wheat straw

33
-

47

22


30

13
-

19

Hemp straw

44
-

45

19
-

21

20
-

22

Rice straw

39

36

10

Bagasse

40

29

13

Beech wood

46

31

23

Fir wood

43

27

29

Poplar wood

50

31

17

27

Conversion of Lignocellulose to
Value
-
added Bioproducts

Plant Biomass

Steam Explosion

Extraction with water

Alkaline extraction

Cellulose

Hydrolysis

(enzymatic or chemical)

Glucose

Energy

Adhesives

Lignin

Hemicellulose

Pentoses (Xylose, Arabinose)

High energy
input needed!

Alternatives
have to be
developed!

e.g.:
Solid State
Fermentation
!

Hydrolysis

(enzymatic or chemical)

Biotechnological
Production

of
Biopolyesters

Development
of efficient
hydrolysis
methods
required
!!

Fermentation to Bioethanol

Petschacher Barbara, Diploma Thesis, Graz University of Technology, 2001

28

Follow
-
up Products of PHAs: Chiral Synthons
for Organic Synthesis

Chiral

synthons
:

Stereoregular

compounds

acting

as

chiral

precursors,

e
.
g
.

production

of

pharmaceuticals,

pheromons,

vitamins,

antibiotics,

aromatics,

perfumes


PHAs
:

Biobased

Polyesters

consisting

mainly

of

optically

pure

monomers

Chiral center

Hydrolysis

leads to a rich source of
bifunctionel,
R(
-
)
-
configurated hydroxy acids.

Market

value

higher

than

for

the

polymer

itself!

Classical Hydrolysis
:

Isolation of PHA

PHA

acidic alcoholysis of the
isolated PHA

Optically pure
monomers

(Seebach et al., 1992; Seebach
and Züger, 1982)

In
-
vivo

degradation

of

PHA

by

adjusting

the

enzymatic

systems

involved

in

intracellular

PHA

metabolism

via

the

cultivation

conditions

(C
-
source,

pH,

T)
;

excretion

of

metabolites

Highly

efficient

process!

App
.

130

PHA

buliding

blocks

reported
-

broad

range

of

possible

chiral

synthons

(Lee et al., 1999)

Process rather complex and highly Solvent
-
demanding!

In
-
vivo

degradation

29

Meat
-

and Bone Meal (MBM) from Slaughterhause
Waste & Rendering Industry


a Precious Nitrogen
Source for Biotechnological Purposes


Classical Utilization of MBM: Animal Feed


Problem: The emerge of
Bovine Spongiform
Encephalopathy

(
BSE
,

Mad Cow Desease

)


Peak: Infection of 3500 head of caddle weekly in
Great Britain


Alternative Utilization: Energy production by
Combustion
→ low value
-
creation


2001:
Task Force Graz University of
Technology

for Safe Utilization of MBM

to
produce value
-
added products!

30

Hydrolysis of Meat
-

and Bone Meal


Precondition of Safe Utilization of MBM:
Hydrolysis of MBM to destroy
prions

Structure of a Prion

Causing BSE

SDS
-
Gel
-
Electrophoresis of alkaline Hydrolysis of MBM

(PhD thesis José Neto, Graz University of Technology, 2006)

Hydrolysis time [h]

31

Production of Meat
-

and Bone Meal

Application of
hydrolyzed MBM for
Biomass production

Possible:

Removal of Lipids prior to
hydrolysis („Degreasing step“)

Application of lipids for

Biodiesel
Production
or as

carbon source for
fermentative Production of e.g.
Biopolymers

32

Concluding Remarks


A

broad

range

of

waste

materials

from

different

origins

exists

to

be

potentially

utilized

for

biotechnological

production

of

biopolymers,

biofuels

and

biochemicals


Selection

of

the

appropriate

waste

stream

for

biotechnological

purposes

depends

on

the

global

region

where

the

production

is

intended
.

Facilities

for

production

should

be

integrated

into

existing

production

lines,

where

the

waste

streams

directly

accrue

(Prime

example
:

Integration

of

sugar
-
,

bioethanol

and

biopolymer

production

in

Brazil)


Improvement

of

upstream

technologies
,

selection

of

optimized

biocatalysts
,

enhanced

downstream

processing

and

autarkic

energy

supply

are

required

to

achieve

cost

efficiency

in

the

production

of

biopolymers,

biofuels

and

biochemicals

from

waste
.

33

Content :


Limitation and rising prices of fossil feedstocks and the increasing
need for

White Biotechnology

: Ecological and Economic needs



Target Areas

: Final products from conversion of waste
materials via

White Biotechnology



What waste materials are available for biotechnological purposes
(occurence and the challenges of their utilization)


Meat and Bone Meal (Slaughterhouses and Rendering
industry)


Sugar Cane industry


Integration of Biofuel and
Biopolymer Production


Whey (Dairy Industry)


Raw Glycerol Liquid Phase (from Biodiesel Production)


Waste Lipids


Cellulosic and Lignocellulosic Feedstocks


Follow
-
up Products of PHAs: Chiral Synthons for Organic
Synthesis



Summary



THANK YOU

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