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Method and application of vacuum freeze-drying Pichia pastoris

A kind of Pichia membranous yeast, vacuum freeze-drying technology, applied in the field of biochemistry

Active Publication Date: 2022-07-05
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Carbohydrates, proteins, and antioxidants are commonly used protective agents that can effectively improve the survival rate of microorganisms, but there are no reports on the application of vacuum freeze-dried Pichia pastoris

Method used

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  • Method and application of vacuum freeze-drying Pichia pastoris
  • Method and application of vacuum freeze-drying Pichia pastoris
  • Method and application of vacuum freeze-drying Pichia pastoris

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Experimental program
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Embodiment Construction

[0010] 1. Preparation of yeast and yeast suspension

[0011] Pichia membranaceus was purchased from China Industrial Microorganism Culture Collection and Management Center. The yeast was preserved in NYDA medium (5 g of yeast extract, 8 g of beef extract, 10 g of glucose, 20 g of agar, and 1000 ml of water) at 4°C. Activate every two months.

[0012] Before the freeze-drying test, Pichia membranaceus was first inoculated on fresh NYDA medium, activated at 28°C for 48 hours, and then the activated yeast was inoculated into NYDB (NYDA without agar) medium. Incubate at 28°C and 180rpm for 24h, then centrifuge at 1351×g and 4°C for 10min, discard the supernatant, add sterile water, and centrifuge again (twice to ensure no residue in the medium), add sterile Water, made 2 x 10 with a hemocytometer 8 CFU / ml yeast suspension.

[0013] 2. Preparation of protective agent and protective agent suspension

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Abstract

The invention relates to a method for vacuum freeze-drying Pichia membranaceus protective agent, which is characterized by comprising the following steps: inoculating Pichia membranaceus on fresh NYDA medium, and after activating for 48 hours, the activated Yeast was inoculated into NYDB (NYDA without agar) medium, incubated at 28°C, 180 rpm for 24 h, and then incubated at 1351× g , Centrifuge at 4°C for 10 min, discard the supernatant, add sterile water, centrifuge again, add sterile water, and use a hemocytometer to make 2 × 10 8 CFU / ml yeast suspension; equal volumes of protectant suspension and yeast suspension were mixed to obtain samples; 1 mL of sample was aliquoted into 10 mL beakers, frozen at −20°C for 12 h, and then stored at a cold trap temperature of Lyophilized at −50°C, vacuum 1–10 Pa.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a method and application of vacuum freeze-drying Pichia pastoris. Background technique [0002] Pichia membranifaciens (Pichia membranifaciens) is a yeast that can effectively control postharvest diseases of fruits and vegetables. disease. However, there are few studies on the commercial application of Pichia membranaceus in the postharvest field. [0003] Vacuum freeze-drying is a seed conservation technique widely used in research and industry. In the food field, this technology is commonly used to produce microbial solid preparations, such as starter, probiotics, biocontrol preparations, etc. Vacuum freeze-drying technology can well meet the requirements of microbial commercialization, but during the freeze-drying process, the survival rate and functional activity of microorganisms such as biocontrol efficacy, acid production capacity, etc. will be affected, especially after fre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04A23B7/155C12R1/84
CPCA23B7/155C12N1/04
Inventor 曾凯芳牛晓影邓丽莉周雅涵姚世响王威浩
Owner SOUTHWEST UNIV