Production of medium-chain-length polyhydroxyalkanoates by sequential feeding of xylose and octanoic acid in engineered Pseudomonas putida KT2440

被引:75
作者
Le Meur, Sylvaine [1 ]
Zinn, Manfred [1 ,2 ]
Egli, Thomas [3 ]
Thoeny-Meyer, Linda [1 ]
Ren, Qun [1 ]
机构
[1] Swiss Fed Labs Mat Sci & Technol Empa, Lab Biomat, CH-9014 St Gallen, Switzerland
[2] HES SO Valais Wallis, CH-1950 Sion, Switzerland
[3] Swiss Fed Inst Aquat Sci & Technol Eawag, CH-8600 Dubendorf, Switzerland
来源
BMC BIOTECHNOLOGY | 2012年 / 12卷
关键词
mcl-PHA; Xylose; Octanoic acid; Pseudomonas putida KT2440; Sequential-feeding; Tailor-made PHA; RECOMBINANT ESCHERICHIA-COLI; BETA-HYDROXYBUTYRIC ACID; POLY(3-HYDROXYBUTYRATE) PRODUCTION; ETHANOL-PRODUCTION; NONANOIC ACID; BATCH; BIOSYNTHESIS; GLUCOSE; POLY(3-HYDROXYALKANOATES); POLY(HYDROXYALKANOATES);
D O I
10.1186/1472-6750-12-53
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: Pseudomonas putida KT2440 is able to synthesize large amounts of medium-chain-length polyhydroxyalkanoates (mcl-PHAs). To reduce the substrate cost, which represents nearly 50% of the total PHA production cost, xylose, a hemicellulose derivate, was tested as the growth carbon source in an engineered P. putida KT2440 strain. Results: The genes encoding xylose isomerase (XylA) and xylulokinase (XylB) from Escherichia coli W3110 were introduced into P. putida KT2440. The recombinant KT2440 exhibited a XylA activity of 1.47 U and a XylB activity of 0.97 U when grown on a defined medium supplemented with xylose. The cells reached a maximum specific growth rate of 0.24 h(-1) and a final cell dry weight (CDW) of 2.5 g L-1 with a maximal yield of 0.5 g CDW g(-1) xylose. Since no mcl-PHA was accumulated from xylose, mcl-PHA production can be controlled by the addition of fatty acids leading to tailor-made PHA compositions. Sequential feeding strategy was applied using xylose as the growth substrate and octanoic acid as the precursor for mcl-PHA production. In this way, up to 20% w w(-1) of mcl-PHA was obtained. A yield of 0.37 g mcl-PHA per g octanoic acid was achieved under the employed conditions. Conclusions: Sequential feeding of relatively cheap carbohydrates and expensive fatty acids is a practical way to achieve more cost-effective mcl-PHA production. This study is the first reported attempt to produce mcl-PHA by using xylose as the growth substrate. Further process optimizations to achieve higher cell density and higher productivity of mcl-PHA should be investigated. These scientific exercises will undoubtedly contribute to the economic feasibility of mcl-PHA production from renewable feedstock.
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页数:12
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