Metabolic engineering of Bacillus subtilis for growth on overflow metabolites

被引:38
|
作者
Kabisch, Johannes [1 ]
Pratzka, Isabel [1 ]
Meyer, Hanna [2 ]
Albrecht, Dirk [3 ]
Lalk, Michael [2 ]
Ehrenreich, Armin [4 ]
Schweder, Thomas [1 ]
机构
[1] Univ Greifswald, Inst Pharm, D-17487 Greifswald, Germany
[2] Univ Greifswald, Inst Biochem, D-17487 Greifswald, Germany
[3] Univ Greifswald, Inst Microbiol, D-17487 Greifswald, Germany
[4] Tech Univ Munich, Dept Microbiol, D-85354 Freising Weihenstephan, Germany
关键词
Bacillus subtilis; Metabolic engineering; Glyoxylate cycle; Expression system; Fed-batch; Acetate; Glycolic acid; HIGH-CELL-DENSITY; ISOCITRATE LYASE; 2,3-BUTANEDIOL; ACETOIN; ACID; CULTIVATION; RESPONSES; SOFTWARE; PROTEINS; VECTORS;
D O I
10.1186/1475-2859-12-72
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: The genome of the important industrial host Bacillus subtilis does not encode the glyoxylate shunt, which is necessary to utilize overflow metabolites, like acetate or acetoin, as carbon source. In this study, the operon encoding the isocitrate lyase (aceB) and malate synthase (aceA) from Bacillus licheniformis was transferred into the chromosome of B. subtilis. The resulting strain was examined in respect to growth characteristics and qualities as an expression host. Results: Our results show that the modified B. subtilis strain is able to grow on the C2 compound acetate. A combined transcript, protein and metabolite analysis indicated a functional expression of the native glyoxylate shunt of B. lichenifomis in B. subtilis. This metabolically engineered strain revealed better growth behavior and an improved activity of an acetoin-controlled expression system. Conclusions: The glyoxylate shunt of B. licheniformis can be functionally transferred to B. subtilis. This novel strain offers improved properties for industrial applications, such as growth on additional carbon sources and a greater robustness towards excess glucose feeding.
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页数:10
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