Title:
MUTANT OF NITRILE HYDRATASE DERIVED FROM CALDALKALIBACILLUS THERMARUM
Document Type and Number:
WIPO Patent Application WO/2021/081868
Kind Code:
A1
Abstract:
Disclosed is a mutant of nitrile hydratase derived from Caldalkalibacillus thermarum, which falls within the technical field of enzyme engineering. The half-life period of the nitrile hydratase mutant Cal.t Nhase-A20V provided in the present invention at 70℃ is about 10 min, the thermal stability of the mutant Cal.t Nhase-A20V has not changed excessively compared with that of a wild enzyme, and the specific enzyme activity of the mutant Cal.t Nhase-A20V is 128％ of that of the wild enzyme. At the same time, the mutant also has better substrate tolerance and product tolerance, the final yield of nicotinamide produced by whole-cell catalysis reaches 598 g/L. The nitrile hydratase mutant Cal.t Nhase-A20V has very good enzymatic properties, and is conducive to future industrial production.
Inventors:
ZHOU ZHEMIN (CN)
LIU ZHONGMEI (CN)
LI TING (CN)
CHEN DEZHI (CN)
ZHANG SAILAN (CN)
ZHOU LI (CN)
CUI WENJING (CN)
CHENG ZHONGYI (CN)
GUO JUNLING (CN)
LIU ZHONGMEI (CN)
LI TING (CN)
CHEN DEZHI (CN)
ZHANG SAILAN (CN)
ZHOU LI (CN)
CUI WENJING (CN)
CHENG ZHONGYI (CN)
GUO JUNLING (CN)
Application Number:
PCT/CN2019/114605
Publication Date:
May 06, 2021
Filing Date:
October 31, 2019
Export Citation:
Assignee:
UNIV JIANGNAN (CN)
International Classes:
C12N1/21; C12N9/88; C12N15/60; C12N15/70; C12P17/12; C12R1/19
Foreign References:
| CN109251882A | 2019-01-22 | |||
| CN109593750A | 2019-04-09 | |||
| CN107177581A | 2017-09-19 | |||
| US20110104690A1 | 2011-05-05 |
Other References:
CHEN JIE; YU HUIMIN; LIU CHANGCHUN; LIU JIE; SHEN ZHONGYAO: "Improving stability of nitrile hydratase by bridging the salt-bridges in specific thermal-sensitive regions", JOURNAL OF BIOTECHNOLOGY, vol. 164, no. 2, 4 February 2013 (2013-02-04), pages 354 - 362, XP028990882, ISSN: 0168-1656, DOI: 10.1016/j.jbiotec.2013.01.021
AKIMASA MIYANAGA, SHINYA FUSHINOBU, KIYOSHI ITO, HIROFUMI SHOUN, TAKAYOSHI WAKAGI: "Mutational and Structural Analysis of Cobalt-Containing Nitrile Hydratase on Substrate and Metal Binding", EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 271, no. 2, 31 December 2004 (2004-12-31), pages 429 - 438, XP055096861, ISSN: 0014-2956, DOI: 10.1046/j.1432-1033.2003.03943.x
SHI YUE,YU HUIMIN,LUO HUI,TIAN ZHUOLING,SHEN ZHONGYAO: "High Level Expression of a Mutated Nitrile Hydratase Gene of Nocardia Sp. in Recombinant Escherichia Coli", JOURNAL OF TSINGHUA UNIVERSITY(SCIENCE AND TECHNOLOGY), vol. 47, no. 12, 31 December 2007 (2007-12-31), pages 2176 - 2179, XP055810021, DOI: 10.16511/j.cnki.qhdxxb.2007.12.008
AKIMASA MIYANAGA, SHINYA FUSHINOBU, KIYOSHI ITO, HIROFUMI SHOUN, TAKAYOSHI WAKAGI: "Mutational and Structural Analysis of Cobalt-Containing Nitrile Hydratase on Substrate and Metal Binding", EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 271, no. 2, 31 December 2004 (2004-12-31), pages 429 - 438, XP055096861, ISSN: 0014-2956, DOI: 10.1046/j.1432-1033.2003.03943.x
SHI YUE,YU HUIMIN,LUO HUI,TIAN ZHUOLING,SHEN ZHONGYAO: "High Level Expression of a Mutated Nitrile Hydratase Gene of Nocardia Sp. in Recombinant Escherichia Coli", JOURNAL OF TSINGHUA UNIVERSITY(SCIENCE AND TECHNOLOGY), vol. 47, no. 12, 31 December 2007 (2007-12-31), pages 2176 - 2179, XP055810021, DOI: 10.16511/j.cnki.qhdxxb.2007.12.008
Attorney, Agent or Firm:
HARBIN SHINEIP INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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