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Kluyveromyces lactis and separation and culture method thereof

A technology of yeast and lactic acid, which is applied in the field of microorganism and tobacco stem processing, can solve the problems of Saccharomyces cerevisiae growth, lack of lactose utilization, etc., and achieve the effect of easy operation, low cost and simple cultivation method

Pending Publication Date: 2020-12-29
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These strains grow well on media with lactose as the carbon source, while S. cerevisiae cannot grow in such media because S. cerevisiae lacks genes related to lactose utilization

Method used

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  • Kluyveromyces lactis and separation and culture method thereof
  • Kluyveromyces lactis and separation and culture method thereof
  • Kluyveromyces lactis and separation and culture method thereof

Examples

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Embodiment 1

[0024] The present embodiment is the method for isolating and cultivating Kluyveromyces lactis KM, comprising the following steps:

[0025] a. Several samples of shrimp paste were collected from Yantai, Shandong, and a single colony was screened out by using YPD solid medium. The single colony was cultured in YPD liquid medium for 12-16 hours, and stored in a -80°C refrigerator;

[0026] b. Select 20 to 30 bacterial colonies from step (a), separate them by streaking in the tobacco leaf solid medium, and cultivate them at 28 to 30°C for 24 to 48 hours;

[0027] c. Select a strain with the best growth from step (b), inoculate it on a YPD solid medium plate, culture it at 28-30° C. for 24-48 hours, and identify it.

[0028] Preparation of YPD solid medium: glucose 20g, peptone 20g, yeast powder 10g, add deionized water to 1000mL, add 15-20g agar to 1000mL liquid medium, sterilize at 121℃ for 15min, cool to about Invert the plates at 50°C, and store at room temperature or in a re...

Embodiment 2

[0033] The morphological, physiological and biochemical properties of the isolated and purified strains were analyzed, and KM genomic DNA was extracted using a fungal genome extraction kit, and 26S rDNA universal primers NL1 (5′-GCATATCAATAAGCGGAGGAAAAG-3′) and NL4 (5′-GGTCCGTGTTTCAA GACGG-3′ ) to amplify the D1 / D2 region sequence of the microorganism and send it to Kunming Shuoqing Biotechnology Company for sequencing. The PCR amplification conditions were: 94°C for 4min; 35 cycles of 94°C for 30s, 50°C for 30s, and 72°C for 60s; 72°C for 10min. The sequence information was submitted to the NCBI database (https: / / www.ncbi.nlm.nih.gov / ) for online comparison analysis.

[0034] The identification results showed that it belonged to Kluyveromyces lactis, and its microbiological classification was named Kluyveromyces lactis KM (Latin name: Kluyveromyces lactis KM).

[0035] The main morphological characteristics and physiological and biochemical properties (see Table 1) of the Kl...

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Abstract

The invention discloses Kluyveromyces lactis. The microbiology classification of the strain is named as Kluyveromyces lactis KM, and the Latin scientific name is Kluyveromyces lactis KM; and the Kluyveromyces lactis is preserved in China Center for Type Culture Collection on September 4, 2020, and the preservation number is CCTCM M2020464. The invention further discloses a method for separating and culturing the Kluyveromyces lactis KM.

Description

technical field [0001] The invention belongs to the technical field of microorganism and cut tobacco stem processing, and in particular relates to Kluyveromyces lactis KM and methods for its separation, cultivation and identification. Background technique [0002] Kluyveromyces lactis is a kind of yeast under the genus Kluyveromyces, and its research began in the late 1950s. Most of the yeast strains obtained at that time were isolated from milk products, such as kumiss, Greek cheese and so on. Vander Walt created the genus Kluyveromyces in 1956, and the name Kluyveromyces lactis was named in honor of Dutch microbiologist Albert Jan Kluyver. [0003] The cell morphology of Kluyveromyces lactis is generally spherical or olive-shaped, which is somewhat similar to that of Saccharomyces cerevisiae. Although there are some differences in the natural habits of strains, the vast majority of strains of K. lactis are isolated from milk products whose main carbon source is lactose. ...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12N1/02C12R1/645
CPCC12N1/02C12N1/16
Inventor 巩效伟朱东来赵伟李廷华李寿波韩熠吴俊张霞洪鎏
Owner CHINA TOBACCO YUNNAN IND
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