Fermentative production of <scp>l</scp>‐glycerol 3‐phosphate utilizing a <i>Saccharomyces cerevisiae</i> strain with an engineered glycerol biosynthetic pathway

Biotechnology and Bioengineering - Tập 100 Số 3 - Trang 497-505 - 2008
Almut Popp1,2,3, Nguyễn Thị Thanh Huyền1,2,3, Kenza-Amel Boulahya4, Carine Bideaux4, Sandrine Alfenore4, Stéphane Guillouet4, Elke Nevoigt1,2,3
1Department of Microbiology and Genetics, Berlin University of Technology, Seestr. 13, D-13353 Berlin, Germany
2fax: 49-30-314-27592
3telephone: 49-30-314-27469
4UMR5504, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, CNRS, INRA, INSA, Toulouse Cedex 4, France

Tóm tắt

AbstractInterest in l‐glycerol 3‐phosphate (l‐G3P) production via microbial fermentation is due to the compound's potential to replace the unstable substrate dihydroxyacetone phosphate (DHAP) in one‐pot enzymatic carbohydrate syntheses. A Saccharomyces cerevisiae strain with deletions in both genes encoding specific l‐G3Pases (GPP1 and GPP2) and multicopy overexpression of l‐glycerol 3‐phosphate dehydrogenase (GPD1) was studied via small‐scale (100 mL) batch fermentations under quasi‐anaerobic conditions. Intracellular accumulation of l‐G3P reached extremely high levels (roughly 200 mM) but thereafter declined. Extracellular l‐G3P was also detected and its concentration continuously increased throughout the fermentation, such that most of the total l‐G3P was found outside the cells as fermentation concluded. Moreover, in spite of the complete elimination of specific l‐G3Pase activity, the strain showed considerable glycerol formation suggesting unspecific dephosphorylation as a mechanism to relieve cells of intracellular l‐G3P accumulation. Up‐scaling the process employed fed‐batch fermentation with repeated glucose feeding, plus an aerobic growth phase followed by an anaerobic product accumulation phase. This produced a final product titer of about 325 mg total l‐G3P per liter of fermentation broth. Biotechnol. Bioeng. 2008;100: 497–505. © 2008 Wiley Periodicals, Inc.

Từ khóa


Tài liệu tham khảo

Alfenore S, 2002, Improving ethanol production and viability of Saccharomyces cerevisiae by a vitamin feeding strategy during fed‐batch process, Appl Microbiol Biotechnol, 60, 67

Arnold WN, 1972, Location of acid phosphatase and ‐fructofuranosidase within yeast cell envelopes, J Bacteriol, 112, 1346, 10.1128/jb.112.3.1346-1352.1972

10.1093/nar/12.20.7721

10.1111/j.1574-6976.2001.tb00570.x

10.1128/AEM.72.3.2134-2140.2006

10.1016/0005-2744(75)90314-9

Cameron DC, 2004, Biobased industrial chemicals, Appl Biochem Biotechnol, 113, 805

10.1385/ABAB:115:1-3:0871

10.1016/j.ymben.2007.03.002

10.1016/0005-2787(78)90276-9

10.1002/anie.200400659

FessnerW.‐D.1997. Enzymatic process for the production of dihydroxyacetone phosphate from glycerophosphate and its use in enzymatic aldol additions patent U.S. patent 5 683 897.

10.1016/j.ymben.2006.06.004

10.1093/nar/30.6.e23

10.1007/BF00260867

Linnemans WA, 1977, Localization of acid phosphatase in Saccharomyces cerevisiae: A clue to cell wall formation, J Bacteriol, 131, 638, 10.1128/jb.131.2.638-644.1977

Lopandic K, 1987, Biochemical and genetic evidence that yeast extracellular protein phosphatase activity is due to acid phosphatase, Biochem Int, 14, 627

10.1016/0014-5793(77)81014-4

10.1016/j.ymben.2004.02.005

10.1002/(SICI)1097-0061(20000330)16:5<463::AID-YEA535>3.0.CO;2-3

10.1074/jbc.272.9.5544

10.1074/jbc.271.23.13875

10.1016/0167-4838(90)90160-H

10.1266/ggs.72.323

10.1128/AEM.68.6.2814-2821.2002

10.1074/jbc.M007164200

10.1093/genetics/149.4.1707

10.1111/j.1432-1033.1991.tb15803.x

10.1016/0076-6879(83)01015-0

Schmidt G, 1963, Acid phosphatase of bakers' yeast: An enzyme of external cell surface, Biochemistry, 2

Schümperli M, 2005, A new approach for the production of DHAP: The system of biotransformations, Kopenhagen: 12th European Congress on Biotechnology.

Schweingruber ME, 1986, Identification and characterization of thiamin repressible acid phosphatase in yeast, J Biol Chem, 261, 15877, 10.1016/S0021-9258(18)66645-1

10.1146/annurev.micro.51.1.285

10.1007/BF01026672

10.1093/nar/17.4.1353

10.1002/yea.320080703

Thông tin
Thông tin xuất bản

Biotechnology and Bioengineering

Tập 100 Số 3

497-505

Thông tin tác giả