Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of fermentation method that improves L-isoleucine yield

A technology of isoleucine yield and isoleucine is applied in the field of fermentation to improve L-isoleucine yield, and can solve the problems of low acid production rate, low sugar-acid conversion rate and the like

Active Publication Date: 2021-09-14
XINTAI JIAHE BIOTECH CO LTD +1
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing L-isoleucine fermentation method still has the problems of low acid production rate and low sugar-acid conversion rate.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of fermentation method that improves L-isoleucine yield
  • A kind of fermentation method that improves L-isoleucine yield
  • A kind of fermentation method that improves L-isoleucine yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Constructed for L- isoleucine-producing strains of engineering: Example 1

[0068] In L- isoleucine producing bacteria C.glutamicumYILW genomic chromosomal DNA as a template, P1 (5'-CGGCTA Gtcgac GTTCAATTGCCATGTCAGT-3 ', as the scribing Sal I restriction site; SEQ ID NO.1) and P2 (5'-GCGCCGA GGATCC GTAATAGGACAACAACGC TC-3 ', as the scribing BamH I restriction site; SEQ ID NO.2) as primers, PCR amplified homoserine dehydratase gene encoding hom. Using BamH I and Sal I and the PCR product was double digested pBR322 plasmid, the ligation product into the chemical transformation E.coli DH5a, cultured on LB plates at a final concentration of 25ug / mL ampicillin 37 ℃. Using primers P3 (5'-GACAATTAATCATCGGCTCG-3 '; SEQ ID NO.3), P4 (5'-CAGGCTGAAA ATCTTCTCTC-3'; SEQ ID NO.4) identified transformants to grow, positive clones, digested verification, sequencing. In L- isoleucine producing bacteria C.glutamicum YILW cloned genomic DNA as template hom gene fragment was connected to dou...

Embodiment 2

[0071] L- isoleucine Production Example 2:

[0072] 1. bacteria expand training:

[0073] Example 1 constructed strains for the species L- isoleucine production were expanded by culturing a seed flask culture and tank culture two seeds, seed was obtained. in:

[0074] A medium for seed culture flask LB liquid medium; seed flask seed culture to a logarithmic phase, the bacteria solution was inoculated to a seed flask cultured seed tank two.

[0075] Media formulations secondary seed culture tank: KH 2 PO 4 1.5g / L, MgSO 4 · 7h 2 O 0.6g / L, (NH 4 ) 2 SO 4 2.25g / L, FeSO 4 · 7h 2 O 10mg / L, MnSO 4 · H 2 O 10mg / L, VB 1 0.3mg / L, glucose 50g / L, yeast extract powder 5g / L, betaine 0.225g / L, lysine 1g / L, methionine 1g / L; pH 7.5 ± 0.1.

[0076] Two seed tank culture conditions: temperature 32 ℃, pH6.8-7.0, 20-30% dissolved oxygen, the tank pressure 0.03MPa, flow rate 15L / min, stirring speed of 400 rpm; the inoculation amount was 10% (volume fraction), culture 10-12h.

[...

Embodiment 3

[0085] L- isoleucine production: Example 3

[0086] 1. bacteria expand training:

[0087] Example 1 strains were constructed by a seed flask cultures were expanded in culture and can be implemented two seeds, seed was obtained. in:

[0088] A medium for seed culture flask LB liquid medium; seed flask seed culture to a logarithmic phase, the bacteria solution was inoculated to a seed flask cultured seed tank two.

[0089] Media formulations secondary seed culture tank: KH 2 PO 4 1.5g / L, MgSO 4 · 7h 2 O 0.6g / L, (NH 4 ) 2 SO 4 2.25g / L, FeSO 4 · 7h 2 O 10mg / L, MnSO 4 · H 2 O 10mg / L, VB 1 0.3mg / L, glucose 50g / L, yeast extract powder 5g / L, betaine 0.225g / L, lysine 1g / L, methionine 1g / L; pH 7.5 ± 0.1.

[0090] Two seed tank culture conditions: temperature 32 ℃, pH6.8-7.0, 20-30% dissolved oxygen, the tank pressure 0.03MPa, flow rate 15L / min, stirring speed of 400 rpm; the inoculation amount was 10% (volume fraction), culture 10-12h.

[0091] 2. Fermentation:

[00...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a fermentation method for increasing the yield of L-isoleucine. The isoleucine production strain is inoculated into a fermentation medium for fermentation. During the fermentation process, dissolved oxygen is controlled and materials are fed step by step to obtain L-isoleucine. ‑Isoleucine. The present invention improves the fermentation yield of L-isoleucine and the sugar-acid conversion rate of the isoleucine-producing bacteria L-isoleucine through fermentation, wherein the yield of L-isoleucine is above 50 g / L, and the sugar-acid conversion rate is above 20%. Compared with the prior art, the yield of L-isoleucine and the conversion rate of sugar and acid are greatly improved. The residual sugar content in the fermentation broth is controlled at 1-2.5% by feeding glucose, which will neither constitute substrate limitation nor cause "glucose effect", and maximize the production capacity of the strain; at the same time, through segmentation Oxygen supply controls the dissolved oxygen in the fermentation broth, which will neither affect the growth of bacteria in the early stage of fermentation nor affect the fermentation in the later stage, so that L-isoleucine can be accumulated in large quantities.

Description

Technical field [0001] The present invention relates to the field of microbial fermentation techniques, and more particularly to a fermentation method of increasing L-isoleucine yield. Background technique [0002] Isoleucine, Ile, also known as "isol", the system is named "α-amino-β-methylprovic acid", chemical C 6 Hide 13 NO 2 A type of aliphatic neutral amino acid. It is one of the eight must be synthesized by people and animals, but must rely on the exogenous supply of one of the extensive supplies, with a variety of physiological functions, which are widely used in the fields of medicine, food, feed and cosmetics. Especially in the pharmaceutical sector, isoleucine can be used to prepare other essential amino acids into composite amino acid infusion, especially high branched chain amino acid infusion, but not only improve the human immunity, but also promote patient rehabilitation and post-recovery. In addition, it is also used in blood-brain barriers, liver prors, chronic c...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/06C12R1/15
CPCC12P13/06
Inventor 岳明瑞熊传波杨明贵杨军崔涛腾义卫
Owner XINTAI JIAHE BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com