Pichia kudriavzevii strain for preparing beta-phenethyl alcohol through biological transformation of L-phenylalanine, culture method and applications thereof

A technology of yeast strain and phenylethanol, which is applied in the field of microorganisms to achieve the effects of optimizing ingredients and culture conditions, increasing production, and high ethanol tolerance

Active Publication Date: 2018-01-26
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problem that natural green β-phenylethanol cannot meet the market demand, and the practical application of increasing the content of β-phenylethanol in traditional brewed food liquor and improving the quality of liquor, th

Method used

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  • Pichia kudriavzevii strain for preparing beta-phenethyl alcohol through biological transformation of L-phenylalanine, culture method and applications thereof
  • Pichia kudriavzevii strain for preparing beta-phenethyl alcohol through biological transformation of L-phenylalanine, culture method and applications thereof
  • Pichia kudriavzevii strain for preparing beta-phenethyl alcohol through biological transformation of L-phenylalanine, culture method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Pichia pastoris Kudri Azwitz ( Pichia kudriavzevii ) Isolation of Y1511

[0048] After pulverizing Gujing Gong Daqu and mixing it evenly, weigh 1 g, add 9 mL of sterile water, oscillate fully and soak for 15 min to make a suspension. Under aseptic conditions, take 0.1 mL of the suspension and dilute it step by step with sterile water to 10 -3 、10 -4 、10 -5 、10 -6 . Take 0.1 mL of each gradient suspension and spread it on the YPD plate, make three parallels for each dilution gradient, and spread sequentially from the low dilution concentration to the high dilution concentration. Place the medium plate at 30 o C cultured in a constant temperature incubator for 2 days to observe the growth of colonies. Pick up a single colony that is spherical, protruding on the surface, milky white, and opaque on the plate, and inoculate it on the YPD plate for multiple times until the bacteria are in the same shape under the microscope. Inoculate a single colony in 50 ...

Embodiment 2

[0049] Example 2 Preservation of Pichia kudriavzevii Y1511

[0050] The Pichia pastoris Kudri Azwitz obtained above ( Pichia kudriavzevii ) Y1511 is saved by the following methods:

[0051] (1) Preservation on slant: Inoculate the purified strains on the YPD slant, place it in the incubator for 48-72 h, and then store it at 4 o C stored in the refrigerator.

[0052] (2) Storage in glycerol tubes: Take 6 mL of 20% glycerol on the purified strain plate, scrape off the colony and mix it with 20% glycerol, divide the mixture into 1.5 mL sterile PE tubes, store at -80 o Save under C.

Embodiment 3

[0053] Example 3 Identification of Pichia kudriavzevii Y1511

[0054] The identification of the above mentioned Pichia kudriavzevii Y1511 includes the following steps:

[0055] Step 1: Morphological Characterization

[0056] In order to identify the yeast strains involved, the following morphological characteristics were observed:

[0057] (1) Colony morphology and cell observation in YPD medium: the pure culture of the strain obtained in Example 1 was inoculated on YPD medium, and its morphological characteristics were observed after 48 h ( figure 1 ), the results are shown in Table 1, and observed under an optical microscope after being magnified 400 times, the results are as follows figure 2 , presenting a long and narrow spindle, a small amount is oval, budding.

[0058] (2) Observation of colony morphology in WL medium: inoculate the screened strains on YPD for activation, culture for 24-48 h and then inoculate into WL medium, 30 o C cultured and observed after 5 da...

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Abstract

The present invention relates to a Pichia kudriavzevii strain for preparing beta-phenethyl alcohol through biological transformation of L-phenylalanine, a culture method and applications thereof, wherein the Pichia kudriavzevii strain is named Pichia kudriavzevii Y1511, is preserved in the China General Microbiological Culture Collection Center on May 31, 2016, and has the preservation number of CGMCC 12567. According to the present invention, the 26S rDNA D1/D2 sequence of the strain and the 26S rDNA D1/D2 sequences of other Pichia kudriavzevii strains have the similarity of 99%; the strain of the present invention has advantages of good sugar tolerance, good ethanol tolerance, good phenethyl alcohol tolerance, wide growth pH valve range and wide temperature range; the screened Pichia kudriavzevii Y1511 of the present invention has the high beta-phenethyl alcohol yield, wherein the detection results show that the beta-phenethyl alcohol yield can achieve 1.0-11.0 g/L, such that the strain can be easily applied in the industrial production of beta-phenethyl alcohol; and the strain of the present invention has advantages of mild reaction conditions, environmentally friendly and natural product, low cost, environmental friendliness and the like.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a newly isolated yeast strain producing β-phenylethanol and its cultivation method and application. Background technique [0002] β-Phenylethyl alcohol is a colorless viscous liquid with a pleasant rose fragrance. The fragrance is soft and elegant, delicate and long-lasting. In the essential oils of many plants such as orchid and lily, especially in the rose essential oil known as "liquid soft gold", its content can reach 60%. The rose fragrance of β-phenylethyl alcohol is very popular among people, and it is the basic component of all rose fragrances, making it widely used in food, cosmetics, daily necessities for washing, tobacco and medicines, etc. The second largest flavor and fragrance. [0003] The global annual consumption of β-phenylethanol is nearly 10,000 tons, which is basically produced by chemical synthesis of chemical raw materials benzene or styrene,...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12P7/22C12R1/84
Inventor 李秀婷滕超范光森孙宝国孙啸涛许岱富志磊
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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