Engineering of thermotolerant bacillus coagulans for production of D(-)-lactic acid

一种基因、乳酸的技术，应用在制造D型乳酸的耐热凝结芽孢杆菌的基因工程领域，能够解决缺乏发酵等问题

Inactive Publication Date: 2014-01-29
UNIV OF FLORIDA RES FOUNDATION INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, despite several attempts to improve the xylose-fermenting properties of these lactic acid bacteria, the lactic acid bacteria used in industry still lack the ability to efficiently ferment five-carbon sugars into lactic acid (25, 31)

Method used

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  • Engineering of thermotolerant bacillus coagulans for production of D(-)-lactic acid
  • Engineering of thermotolerant bacillus coagulans for production of D(-)-lactic acid
  • Engineering of thermotolerant bacillus coagulans for production of D(-)-lactic acid

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Embodiment 1

[0094] Example 1 - Construction of Bacillus coagulans that produces D-lactic acid and is resistant to heat

[0095] Raw materials and methods

[0096] Strains and plasmids

[0097] The prior art has been described for the Bacillus coagulans wild-type strain P4-102B (27). Escherichia coli strain Top10 (Invitrogen) and Bacillus subtilis strain HB1000 (11) were used as hosts during the construction of the various plasmids used in this study. Plasmid pGK12 carries chloramphenicol and erythromycin resistance genes and replicates in several Gram-positive bacteria and E. coli (17, 21). Although this plasmid has a broad host range, its replication is inherently restricted to temperature conditions ≤42°C. Due to the temperature-sensitive nature of plasmid pGK12 replication at 50°C, this provides an opportunity to screen for Bacillus coagulans with chromosomal DNA insertions that can grow at temperatures ranging from 50°C to 55°C. Plasmid pGK12 and its derivatives were maintaine...

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Abstract

Genetically modified microorganisms having the ability to produce D(−)-lactic acid at temperatures between 30° C. and 55° C. are provided. In various embodiments, the microorganisms may have the chromosomal lactate dehydrogenase (ldh) gene and / or the chromosomal acetolactate synthase (alsS) gene inactivated. Exemplary microorganisms for use in the disclosed methods are Bacillus spp., such as Bacillus coagulans.

Description

[0001] Cross References to Related Applications [0002] This application claims the rights of U.S. Provisional Patent Application Serial No. 61 / 416002 filed on November 22, 2010, and the disclosure content of this patent is hereby fully incorporated into this application by way of data reference, including all drawings, tables and nucleic acid sequence. [0003] governmental support [0004] This invention was made with government support under DOE Grant No. DE-FG36-04GO14019. The government has certain rights in this invention. Background technique [0005] Petroleum is not only a major source of fuel, but also a feedstock for the manufacture of many polymers used by the plastics industry. The limited reserves of petroleum, coupled with the negative environmental impact of its use, have led to an increasing focus on alternative, renewable fuel sources and compounds that could replace petroleum (23). Fermentation of carbohydrates has been shown to produce a variety of sh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74C12N15/52C12P7/56C12R1/07
CPCC12N9/1022C12P7/56C12N9/0006C12N1/20C12N15/52C12N15/75
Inventor 王庆昭基尔纳升·T·尚穆根朗尼·欧尼尔·英格拉姆
Owner UNIV OF FLORIDA RES FOUNDATION INC
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