Leibniz-Institute for Agricultural Engineering and Bioeconomy (ATB)

Max-Eyth-Allee 100, D-14469 Potsdam, GERMANY

Fon: +49(331)5699-852, email: jvenus@atb-potsdam.de

http://de.linkedin.com/pub/joachim-venus/15/276/3b2/

Fermentation and

purification of

lactic acid using

membrane

processes

1927 Experimental farm of the Agricultural University Berlin

1933 Independent research center on agricultural mechanization

1952 Central institute of agricultural engineering of East Germany

1992 Reestablished after the reunification of Germany

Today: Leibniz Institute for Agricultural Engineering and Bioeconomy

- member of the Leibniz Association

History Our mission:

Our research is aimed at sustainable

intensification. We analyze, model and evaluate

bio-economic production systems. We develop and

integrate new technologies and management

strategies for a knowledge-based, site-specific

production of biomass, and its use for food, as bio-

based materials and fuels - from basic research to

application.

The National Research

Strategy Bioeconomy 2030

was published under the

direction of the Federal

Ministry of Education and

Research (BMBF),

together with six additional

ministries.

the strategy involves

five key fields of

action

Technology

assessment in

agricultural

systems

Technologies and processes for crop

production and livestock management

• Ensuring sustainable agricultural production

R & D at ATB in the direction of BIOECONOMY

Cascading use of biomass / Biorefinery concepts

Research Program

„Material and energetic use of biomass “

Coordination: Dr. Joachim Venus

Biomass provision

(Cultivation, harvest, storage… e.g. short

rotation wood, hemp)

Material use

(Fibers, insulation, biotechnological

products)

Energetic use (Biogas, wood pellets,

biochar)

Consideration of the entire value chain –

System‘s approach

Valorization of residues, sidestreams etc.

19.10.2017 5

Industrial Biotechnology - Using renewable resources for industry

Biobased products and processes from renewable resources not only help

preserve the environment and climate,

but also make a significant contribution to the structural change from a

petrochemical to a biobased industry, with related opportunities for growth

and employment. Industrial biotechnology, also known as white

biotechnology, is an important driving force in this transition.

March 2014 May 2012 2010/2011_en

19.10.2017 6

19.10.2017 7

International Journal of Biological Macromolecules

Volume 98, May 2017, Pages 447–458

Review Biotransformation of lignocellulosic

materials into value-added products—A review

M. Bilal, M. Asgher, H.M.N. Iqbal, H. Hu, X.

Zhang

Building blocks that could be produced via fermentation

Numbers next to biochemicals designate the total annual production in thousands of t

SpecialChem - Aug 2014 - http://www.specialchem4bio.com/news/2014/08/20/lactic-acid-market-estimated-to-reach-usd-3577-5-mn-by-2019-

marketsandmarkets

The market for lactic acid is growing as it is largely used in various industrial applications such as in biodegradable polymers,

food & beverages, personal care products, and pharmaceutical industries. The lactic acid market is mainly driven by its end-

use industries.

In 2013, Biodegradable polymers formed the largest application for lactic acid, followed by food and

beverages. The lactic acid market is estimated to grow at a CAGR of 18.8% from 2014 to reach $3,577.5

million by 2019.

The processes for producing lactic acid from biomass/residues

include the following 4 main steps:

19.10.2017 9

(1) Pretreatment - breaking down the structure of the feedstock matrix

(2) Enzymatic hydrolysis - depolymerizing biopolymers like starch, cellulose etc.

to fermentative sugars, such as glucose (C6) and xylose (C5), by means of

hydrolytic enzymes

(3) Fermentation - metabolizing the sugars to lactic acid, generally by LAB

(4) Separation and purification of lactic acid - purification of lactic acid to meet

the standards of commercial applications

Pilot plant facility for lactic acid fermentation at Leibniz-Institute for Agricultural Engineering Potsdam-Bornim / ATB

19.10.2017 10

Biorefinery-concept for (1st, 2nd, 3rd…?) biomass feedstocks

- BIOCONVERSION -

Pleissner, D.; Dietz, D.; van Duuren, J.B.J.H.; Wittmann, C.; Yang, X.; Lin, C.S.K.; Venus, J.: Biotechnological production of organic acids from renewable resources. Advances in Biochemical Engineering/Biotechnology, pp 1-38, 07 March 2017, http://link.springer.com/chapter/10.1007/10_2016_73

(Different) Composition & Behaviour

of (lignocellulosic) Biomass

19.10.2017 11

M.A. Abdel-Rahman et al.

Journal of Biotechnology 156 (2011) 286– 301

V. Menon, M. Rao

Progress in Energy and Combustion

Science 38 (2012) 522-550

N. Mosier et al. / Bioresource Technology 96 (2005) 673–686

19.10.2017 12

Table 1: Overview of chemicals that are currently

produced, or could be produced, from biomass

together with their respective market type, size of

the market, and potential biomass feedstock.

Major players involved are also given.

M.A. Abdel-

Rahman et al.

Journal of

Biotechnology

156 (2011) 286–

301

Chemicals from Biomass: A Market Assessment of

Bioproducts with Near-Term Potential Mary J. Biddy, Christopher Scarlata, and Christopher Kinchin - National Renewable Energy Laboratory

19.10.2017 13

Data Gaps

Scale-up of lactic acid production would require clean, cheap sugars

from lignocellulosic biomass to compete with commodity sugar and

starch substrates. There is a lack of data about lactic acid production

and purification from biomass hydrolysates, including issues of C5

sugar utilization, although it appears work has started to address some of

these issues.

This report is available at no cost from the National Renewable Energy Laboratory

(NREL) at www.nrel.gov/publications

Technical Report NREL/TP-5100-65509 - March 2016/Contract No. DE-AC36-08GO28308

19.10.2017 14

Starchy materials (cereals, industrial grade corn/potatoe starch, tapioca)

Green biomass (alfalfa, grass juice, lupine, sweet sorghum, forage rye, silage, coco juice)

Lignocellulosics (wood/straw hydrolysates, 2ndG sugars, bagasse)

Residues & By-products (oilseed cake/meal, thick juice, molasses, whey, coffee residues, waste bread, waffle

residues, algae biomass, fruit residues, rice bran, meat & bone meal, OMSW…)

tapioca

bagasse

waste bread

pine

coco juice

2G sugars 2G sugars

1G/2G sugars

green biomass

several residues…

lupine

cereals,

straw

sorghum

Fermentation feedstocks already tested:

silage

algae

biomass

Coffee residues,

cassava, bagasse

Pleissner, D.; Venus, J.: Agricultural residues as feedstocks for lactic acid fermentation. - ACS Symposium Series, Vol. 1186 "Green Technologies for the Environment" (2014) pp 247–263

rice bran

Olive mill

solid waste

bagasse

19.10.2017 15

Pro

cess s

tep

s f

or t

he m

an

ufa

ctu

re o

f la

cti

c a

cid

Raw material storage

5 m3 (Trevira®UV–silo, HIMEL

Maschinen GmbH & Co. KG)

Hydrolysis 1 m³ stirred vessel; 0.5 m³ storage

tank (Apparate und Behältertechnik

Heldrungen Gmbh)

Pre-, Microfiltration

0.8 mm coarse filter (Sommer &

Strassburger GmbH & Co. KG),

Microfiltration (ZrO2-TiO2

CeRAM®INSIDE, TAMI Industries France)

Sterilization of the

nutrient broth

2 x 400 L, 2 x 250 L stirred vessels

(Apparate und

Behältertechnik Heldrungen Gmbh)

Fermentation with

cell retention

Pilot fermentor Type P, 450 L

(Bioengineering AG)

MOLSEP®Hollow fibre PES membrane

(FS10-FC-FUS50E2, MICRODYN-NADIR

GmbH/Daicen Membrane Systems

Ltd.)

Venus, J.: Res. J. Biotech 2009, 4, No. 2, 15-22

19.10.2017 16

The Copenhagen Declaration

for a Bioeconomy in Action

…

9. The conference also underlined the

need for new pilot and demonstration

plants and scaling up facilities, in

particularly biorefineries. It was stressed,

that the development of these facilities requires

smart integration of various funding sources,

including the Common Agricultural Policy, the

Common Fisheries Policy, the Cohesion Policy,

the Renewable Energy Policy, Horizon 2020, and

private investments.

…

Copenhagen conference “Bioeconomy in Action” on 26 March - 28 March 2012

Universities, Research Institutes, SMEs

Applied & basic research

Industry Industrial application

Large-scale production

„Valle

y o

f death

“

Carus/Carrez/Kaeb/Ravenstijn/Venus: Level Playing Field for Bio-based Chemistry

and Materials. – bioplastics MAGAZINE [03/11] Vol. 6, 52-55

Scale-up of bioprocesses

19.10.2017 18

Pilot plant facility • pilot facility for production of lactic acid at the ATB consequently fills a gap in the

various phases of bioprocess engineering

• provision of product samples is intended to open up

the possibility of interesting partners in industry with

specific product requirements in various applications

BIOSTAT® Bplus (Sartorius BBI Systems GmbH, Germany)

equipped with a digital control unit DCU for the

continuous fermentation with cell recycling

scale up

Pilot fermentor Type P, 450 L (Bioengineering AG)

Venus, J.; Richter, K.: Eng. Life Sci. 2007, 7, No. 4, 395-402

Venus, J.: Feedstocks and (Bio)Technologies for Biorefineries. – In: G.E. Zaikov, F. Pudel, G. Spychalski (Eds.), Renewable

Resources and Biotechnology for Material Applications (pp. 299-309), Nova Science Publishers, 2011 (ISBN: 978-1-61209-521-9)

19

Continuous fermentation process with cell retention

„…Unbelievably, a cheap product such as lactic acid

has until recent-ly been manufactured by batch

processes, although continuous production is the only

economically viable way of producing it…”

Venus, J.: Res. J. Biotech 2009, 4, No. 2, 15-22

0

20

40

60

80

100

0 100 200 300 400 500 600 700

time [hours]

biomass

lactate

0

20

40

60

80

100

0 100 200 300 400 500 600 700

time [hours]

biomass

lactate

20

Pleissner, D.; Qi, Q.; Gao, C.; Perez Rivero, C.; Webb,

C.; Lin, C.S.K.; Venus, J.: Valorization of organic

residues for the production of added value chemicals: A

contribution to the bio-based economy. Biochemical

Engineering Journal 116 (2016) 3-16

Continuous mode

fermentation with cell

retention by hollow fibre

membranes

Example coffee residues: residues from the coffee production

Mass balance from coffee pulp to lactic acid (*downstream

processing was not optimized). All figures are based on dry

weight.

Pleissner, D.; Neu, A.-K.; Mehlmann, K.; Schneider, R.; Puerta-

Quintero, G.I.; Venus, J.: Fermentative lactic acid production from coffee

pulp hydrolysate using Bacillus coagulans at laboratory and pilot scales.

Bioresource Technology 218 (2016) 167–173

19.10.2017 22

Example agro-residues: Sugarcane bagasse

http://www.unicamp.br/unicamp/ju/

540/conversao-de-bagaco-da-

cana-abre-frente-para-producao-

de-polimero-verde

„ENZYMES FOR LIGNOCELLULOSIC FEEDSTOCK DEGRADATION AND PRODUCTION OF SUGARS (C6, C5) AND VALUE ADDED PRODUCTS“

(2010-1.1-203-070-026, 2010 - 2013)

BMBF / DLR - FKZ RUS 10/128

23

A.N.Bach Institute of Biochemistry of Russian Academy of

Sciences, INBI

www.inbi.ras.ru

Leninsky Prospect, 33-2

119071 Moscow

Targets of Russian Partner (INBI):

Enzymes for lignocellulosic feedstocks

degradation and production of sugars (C6,

C5) and value added products

Targets of German Partner (ATB):

Conversion of simple sugars (C6, C5) to

added value products – fuels, solvents,

organic acids, biopolymers, misc. organics

Leibniz-Institut für Agrartechnik Potsdam-Bornim e.V., ATB

www.atb-potsdam.de

Max-Eyth-Allee 100

D-14469 Potsdam

0

10

20

30

40

50

60

st 162

st 306

st 312

st A27

st A40

st A81

st A83

st A116

st A120

st A122

st A123

concentr

ati

on [

g/L

]

st(rain) screening

Lactate [g/L] Glucose [g/L] Xylose [g/L]

Glaser, R.; Venus, J.: Screening of Bacillus coagulans strains in

lignin supplemented minimal medium with high throughput

turbidity measurements. Biotechnology Reports 4 (2014) 60–65.

DOI: 10.1016/j.btre.2014.08.001

Example green biomass: Grass processing with a screw

press into juice and pellet

19.10.2017 24

Vodnar, D.C.; Venus, J.; Schneider, R.; Socaciu, C.: Lactic Acid Production

by Lactobacillus paracasei 168 in Discontinuous Fermentation Using Lucerne

Green Juice as Nutrient Substitute. Chemical Engineering & Technology

33(2010) No. 3, 468-474

Papendiek, F.; Venus, J.: Cultivation and fractionation of leguminous biomass

for lactic acid production. Chem. Biochem. Eng. Q., 28 (3) 375–382 (2014)

0

6

12

18

0 10 20 30 40 50time [hours]

Bio

mass

0

50

100

150

Glu

co

se

, L

acta

te

Typical time course of a batch lactic acid

fermentation supplemented by conventional nutrients

(open symbols), and green juice (solid symbols) as an

alternative source

[g·L-1]Glucose[g·L-1]Biomasse

[g·L-1]Lactat

[g·L-1]Glucose[g·L-1]Biomasse

[g·L-1]Lactat

A cluster of 17 partners will explore the recycling of

vegetable raw materials along the complete value chain

Start of project: July 1st, 2009

2nd phase: July 1st, 2012 – June 30th, 2014

http://www.synrg-cluster.de/index.html

WP2: Technologies and Processes for the harvest, pre-treatment, and

purification, in particular: utilization of (oilseed) residues

Example agri-residues: Rapeseed cake/meal

Venus, J.: Chemie Ingenieur Technik, Volume 86, Issue 9, Sept. 2014, Pages: 1603–1604

ProcessNet-Jahrestagung und 31. DECHEMA-Jahrestagung, 30. September – 2. Oktober 2014

0

10

20

30

40

50

60

70

80

0 10 20 30 40 50 60 70time [hours]

conce

ntr

ati

on [

g/L]

19.10.2017 26

Example wheat straw: Sugar uptake &

product formation

Lactate [g·L-1

]

Sugars [g·L-1

]

Fermentation ended after 50-60 hours with a yield

of nearly 100% and 64 g/L (top left)

(Total) Sugars (firstly Glucose followed by

Arabinose/Xylose with residues of Disaccharides)

have been used completely in the same time

(bottom left)

(Max) Lactate productivity (>5 gL-1h-1) is much

higher than comparable published results

[Li/Cui: Microbial Lactic Acid Production from Renewable

Resources, pp. 211-228. In O.V. Singh and S.P. Harvey

(Eds.), Sustainable Biotechnology - Sources of Renewable

Energy. Springer, 2010]

WO 2013164423 A1; WO 2013164425 A1

Pleissner, D.; Venus, J.: Agricultural residues as feedstocks for lactic acid fermentation. - ACS

Books "Green Technologies for the Environment" (2014) Chapter 13, pp 247–263

0

5

10

15

20

25

30

35

0 10 20 30 40 50 60 70time [hours]

Dis

acc

h,

Ara

, Xyl

[g/L]

0

10

20

30

40

50

60

70

Glu

, T

ota

l su

gars

[g/L]

DisacchAra

XylGluTotal

Example food waste: Bakery industry

González, R.; Venus, J.: BREA4PLA Project. V Intern. Seminar “Biopolymers & Sustainable Composites”, AIMPLAS (6&7 March, 2014 in Valencia)

Venus, J.: Utilization of Waste Bread for Lactic Acid Fermentation. ASABE and CSBE | SCGAB Annual International Meeting, July 13-16, 2014 – Montréal,

Volume 1, 2014, 557-562

& Partner

The award ceremony for the best 2015 Life projects took place in the context of "Green Week" (30 May to 3 June) in the Egg Conference Centre in Brussels

Ongoing projects

ProFIT „Polymerisierbare

Milchsäure aus alternativen

Zuckern“, Antrags-Nr.:

80170179

national: EFRE

(FuE) / ILB

2017-

2020

Chemical building blocks from

versatile MSW

biorefinery — PERCAL

EU/BBI, Grant

agreement No:

745828

2017-

2020

http://percal.aimplas.es/

HyAlt4Chem “Säurebasierte

Hydrolyse von unbehandelten

Altholzrecyclaten zur

Bereitstellung von

Biochemikalien”

national: BMBF/

VDINDE Innovation

+Technik GmbH;

FKZ: 03VNE1032D

2017-

2020

https://www.bmbf.de/de/8-

millionen-tonnen-

gebrauchtes-holz-koennten-

wiederverwendet-werden-

4547.html

19.10.2017 28

Fermentation with

cell retention

Pilot fermentor Type P, 450 L

(Bioengineering AG)

MOLSEP®Hollow fibre PES membrane

(FS10-FC-FUS50E2, MICRODYN-NADIR

GmbH/Daicen Membrane Systems

Ltd.)

Softening 2 x 135 L PUROLITE, 1,5 m³/h (UIT

GmbH Dresden)

Monopolar/Bipolar

Electrodialysis

FT–EDR/ ED4–15; 7,68 m2

monopolar/3,2 m2 bipolar (FuMA-Tech

GmbH Vaihingen)

Ion exchange Cationic resin, 50 L; Anionic resin,

2 x 90 L (UIT GmbH Dresden)

Decolorization Activated carbon, several specific

resins

Evaporation

«chemReactor» CR15 (Büchi AG

Uster/Switzerland);

Rotary Evaporator LABOROTA 20 S

(Heidolph Instruments)

29

Dow

nst

ream

pro

cess

ing o

f ra

w lacta

tes

& lacti

c a

cid

19.10.2017 30

Effect of several down-streaming steps on

the purity of lactic acid

Na-Lactate (Fermentation)

down-stream processing (Filtration, Softening, Electrodialysis,

Ion exchange, Evaporation)

lacticacid

PO4-P

Cl-

SO42-

Nges

Na+

K+

Mg2+

Ca2+

lacticacid

PO4-P

Cl-

SO42-

Nges

Na+

K+

Mg2+

Ca2+

0

500

1.000

1.500

2.000

2.500

3.000

3.500

4.000

4.500

5.000

Start_Ferm End_Ferm M F Soft EDR EXA133 EXC08 Evap

ion

s [m

g/L

]

0

100

200

300

400

500

600

700

800

900

1000

lacta

te [

g/L

]

Raw material storage 5 m3 (Trevira®UV–silo, HIMEL Maschinen GmbH & Co. KG)

Hydrolysis 1 m³ stirred vessel; 0.5 m³ storage tank (Apparate und Behältertechnik

Heldrungen Gmbh)

Pre-, Microfiltration

0.8 mm coarse filter (Sommer & Strassburger GmbH & Co. KG),

Microfiltration (ZrO2-TiO2 CeRAM®INSIDE, TAMI Industries France),

0.3 µm

Sterilization of the nutrient

broth/additives

2 x 400 L, 2 x 250 L stirred vessels (Apparate und

Behältertechnik Heldrungen Gmbh)

Fermentation with cell

retention

Pilot fermentor Type P, 450 L (Bioengineering AG)

MOLSEP®Hollow fibre PES membrane (FS10-FC-FUS50E2, MICRODYN-

NADIR GmbH/Daicen Membrane Systems Ltd.)

Softening 2 x 135 L PUROLITE, 1,5 m³/h (UIT GmbH Dresden)

Monopolare/Bipolare

Electrodialysis

10 cat- and 10 anions exchange membrans unit Type 100 (Deukum,

Germany)

Ion exchange Cationic resin, 50 L; Anionic resin, 2 x 75 L

(UIT GmbH Dresden, Germany)

Process steps for the manufacture of lactic acid

31

Scale-up of fermentation processes challenging.

Production of inexpensive compounds in continuous flow cultures reasonable.

Cell retention in continuous flow cultures significantly increases volumetric productivity.

Conventional downstream processing including filtration, dialysis and chromatography appropriate.

Pure lactic acid formulation can be obtained even with complex substrates.

32

Conclusions

0

500

1.000

1.500

2.000

2.500

3.000

3.500

4.000

4.500

5.000

Start_Ferm End_Ferm M F Soft EDR EXA133 EXC08 Evap

ion

s [m

g/L

]

0

100

200

300

400

500

600

700

800

900

1000

lacta

te [

g/L

]

0

2

4

6

8

10

12

14

16

0 10 20 30 40 50 60 70

Biomass [g/L]

Pro

duct

ivit

y [g

·L-1

·h-1

]

3 L

250 L

Thank you for your attention!

More information: www.atb-potsdam.de

ATB 2016

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