Signal peptide, L-glutamic acid synthesized using konjac flour and methods of using same

Patent metadata

Field	Value
Publication number	US-10351816-B2
Application number	US-201515736544-A
Country	US
Kind code	B2
Filing date	Nov 23, 2015
Priority date	Nov 23, 2015
Publication date	Jul 16, 2019
Grant date	Jul 16, 2019

Abstract

The present invention relates to application of a novel signal peptide in L-arginine and its derivatives production from konjac powder, which belongs to the field of gene engineering, enzyme engineering and metabolism engineering. The present invention fused the signal peptide set forth in SEQ ID NO.1 with the β-mannanase of Bacillus subtilis CCTCC M 209200, and expressed the fused gene in the strain with high L-arginine yield. The recombinant strain Corynebacterium crenatum CGMCC 0890/p MSPman had advantages on utilizing cheaper konjac powder as substrate, and after fermenting for 96 hours in a 5 L bioreactor, the L-arginine yield reached 45 g/L. Another two recombinant strains were constructed based on Corynebacterium crenatum CGMCC 0890/pMSPman, and after fermenting for 96 hours in a 5 L bioreactor, the L-ornithine yield and L-citrulline reached 23.5 g/L and 26.3 g/L respectively.

First claim

The invention claimed is:

1. A mannase signal peptide that is encoded by a non-naturally occurring nucleotide sequence set forth in SEQ ID NO: 1, wherein the mannase signal peptide mediates secretion of beta-mannase.

Assignees

• Univ Jiangnan

Inventors

• Rao Zhiming
• Xu Zhenghong
• Zheng Junxian
• Xu Meijuan
• Yang Taowei
• Zhang Xian

Classifications

• C12Y302/01025

  Beta-mannosidase (3.2.1.25), i.e. mannanase · CPC title

• C07K14/32

  from Bacillus (G) · CPC title

• C12P13/10 Primary

  Citrulline; Arginine; Ornithine · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title