Screening and application of L-proline high-producing Brevundimonas sp. (JNPP-1)

A technology of JNPP-1 and Monomonas, which is applied in the field of screening and application of Brevimonas deficient JNPP-1 to achieve the effect of high transformation rate and high concentration

Active Publication Date: 2011-07-20
JIANGNAN UNIV
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  • Claims
  • Application Information

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  • Screening and application of L-proline high-producing Brevundimonas sp. (JNPP-1)
  • Screening and application of L-proline high-producing Brevundimonas sp. (JNPP-1)

Examples

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Embodiment 1

[0027] Example 1 Isolation and Identification of Brevundimonas JNPP-1 and Preservation of the Strain

[0028] (1) Isolation and screening of Brevundimonas JNPP-1

[0029] The present invention obtains soil samples from domestic chemical plants, weighs 1.0-2.0g soil samples in a 250mL Erlenmeyer flask, suspends them with 10-50mL sterile water, then places them in a water bath, treats them at 60°C for 10min, and performs gradient dilution after cooling , take 0.2mL of the diluted solution and spread it on a broth solid plate, and after incubating at 30-40°C for 24-36h, pick a single colony and perform an oxidase test. The strains with positive oxidase reaction were selected for fermentation verification. Pick a single colony of the strain with positive oxidase reaction and inoculate it into a fresh broth liquid medium for 12 hours, then inoculate the cell liquid culture with 5% (w / w) inoculum in a 250mL triangle containing 30mL fermentation medium In the bottle, 30-40°C, shaki...

Embodiment 2

[0034] Example 2 Production of L-proline by Fermentation of Brevundimonas

[0035] (1) Preparation of cell liquid culture of Brevundimonas diminuta JNPP-1 strain

[0036]Pick a ring of Brevundimonas diminuta JNPP-1 strain on the solid broth medium, inoculate it in a 250mL Erlenmeyer flask filled with 50mL LB medium, and place it on a shaker under the condition of 30°C-40°C. Cultivate at a rotational speed of 200r / min for 9-12h to the mid-logarithmic growth phase, that is, a cell liquid culture of Brevundimonas diminuta JNPP-1 strain is prepared.

[0037] (2) The composition and ratio of the fermentation medium are: carbon source 50-150g / L; organic nitrogen source 6-30g / L; (NH 4 ) 2 SO 4 20-30g / L; K 2 HPO 4 0.1-5.0g / L; MgSO 4 0.1-5g / L; ZnSO 4 0.01-0.10g / L; MnSO 4 0.01-0.10g / L; sodium glutamate 40-60g / L; biotin 50-200μg / L; thiamine 50-200μg / L; pH 6.5-7.4, sterilized under high pressure steam at 115℃ for 20min. The carbon source is selected from one or two of glucose,...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to screening and application of an L-proline high-producing Brevundimonas sp. JNPP-1. An L-proline high-producing pseudomonad JNPP-1 is screened out for the first time; and by combining morphological characteristics and physiological biochemical characteristics, the strain JNPP-1 is identified as a Brevundimonas sp. The strain is preserved at China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC NO.3828. The strain can produce glucoses into L-proline, and the yield can be up to 32.67g / L. The L-proline produced by the strain is one of important amino acids for synthesizing human proteins, is an important material for amino acid transfusion, and has been widely used in multiple fields, such as food, medicine and other industries.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the screening and application of Brevundimonas sp. JNPP-1, which produces high L-proline. Background technique [0002] L-proline is widely used in medicine, agriculture, chemical industry and food. It is mainly used as amino acid medicine in medicine. It is one of the raw materials of compound amino acid infusion. It is used for malnutrition, protein deficiency, severe gastrointestinal tract Protein supplementation after illness, burns and surgery. It is also the raw material for the preparation of Captopril, a drug for treating refractory hypertension and congestive heart failure. In the synthesis industry, L-proline can participate in the induction of asymmetric reactions, and can be used as a catalyst for reactions such as hydrogenation, polymerization, and water-mediated reactions. It has the characteristics of strong activity and good stereospecificity; because L-p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/24
Inventor 许正宏窦文芳张晓梅许泓瑜陆震鸣赵世杰陈敬华史劲松
Owner JIANGNAN UNIV
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