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CN-121628785-B - Pichia kluyveri, pediococcus acidilactici and application of co-fermentation of pichia kluyveri and pediococcus acidilactici in degumming of fresh coffee beans

CN121628785BCN 121628785 BCN121628785 BCN 121628785BCN-121628785-B

Abstract

The invention discloses a Kluyveromyces, a Pediococcus acidilactici and application thereof in degumming fresh coffee beans, belongs to the technical field of microbial fermentation, and can be used for jointly fermenting and applying two strains (KI 23 and LII 35) to degumming the fresh coffee beans and also can be used for preparing a biological degumming preparation of the coffee. When the compounding ratio of the strain compound bacteria liquid is KI 23:LII35=2:1, the fermentation temperature is 32 ℃, the concentration is 10 7 CFU/mL, and the fermentation time is 36h, the pectase activity can reach 982.985u/g, pectin can be effectively decomposed, the biological degumming of fresh coffee beans is realized, and the application prospect is good.

Inventors

  • HU YONGJIN
  • ZHOU YONGXIU
  • NI JIE
  • LI HONG
  • LU KAIHUA

Assignees

  • 云南农业大学

Dates

Publication Date
20260512
Application Date
20260202

Claims (9)

  1. 1. A coffee bean degumming agent for fermentation degumming is a mixed seed bacterial suspension of Pichia pastoris (Pichia kluyveri) LII35 and Pediococcus acidilactici (Pediococcus acidilactici) KI23, wherein the Pichia pastoris LII35 is preserved in China general microbiological culture Collection center (CGMCC) No. 35115 at 7.7.2025, the preservation address is North Star Xidelu No.1 and No. 3 in the Beijing area, the Pediococcus acidilactici KI23 is preserved in China general microbiological culture collection center at 7.7.2025, the preservation number is CGMCC No. 35114, and the preservation address is North Star Xidelu No.1 and No. 3 in the Beijing area.
  2. 2. The degumming agent for fermented degummed coffee beans according to claim 1, wherein the mixing ratio of pichia kluyveri LII35 and pediococcus acidilactici KI23 in the mixed seed bacterial suspension is (3-1): 1-3.
  3. 3. A degumming agent for fermented degummed coffee beans according to claim 1, characterized in that the mixed seed bacterial suspension has a bacterial liquid concentration of 10 5 -10 9 CFU/mL.
  4. 4. A method of degumming a coffee bean using the degumming agent of claim 1, comprising the steps of: 1) Screening and cleaning, namely pouring fresh coffee fruits into a water tank after removing unripe, overripe, fermented, worm-eaten and mildewed defective fruits, injecting clear water, stirring and fishing out floaters; 2) Peeling the fresh coffee fruits obtained in the step 1) by using a peeling machine to obtain fresh coffee beans with pectin layers; 3) Adding fresh coffee beans with fruit gum layers into a fermentation tank, adding a coffee bean degumming agent compounded by pichia kluyveri LII35 and Pediococcus acidilactici KI23, regulating pH by adding HCl or NaOH, decomposing pectin by utilizing the action of microorganisms, fermenting at 20-36 ℃ for 12-60 hours to gradually separate the pectin layer from the coffee beans; 4) Stirring and drying, namely spreading the coffee beans on a airing frame and placing the coffee beans in a climate simulation box after fermentation is completed, setting the temperature of the simulation box to be 22-36 ℃ and the humidity of the simulation box to be 40-50%, and stirring the coffee beans at fixed time until the moisture content is reduced to 10-12%, so as to obtain dried coffee beans; 5) Dehulling, namely removing parchment by using a dehulling machine to obtain roasted green coffee beans.
  5. 5. The method of claim 4, wherein the ratio of the pichia kluyveri LII35 to the pediococcus acidilactici KI23 in step 3) is 1:2.
  6. 6. The method according to claim 4, wherein the concentration of the compounded bacterial liquid in the step 3) is 10 7 CFU/mL.
  7. 7. The method according to claim 4, wherein the fermentation time in step 3) is 36h.
  8. 8. The method according to claim 4, wherein the fermentation temperature in step 3) is 32 ℃.
  9. 9. The method according to claim 4, wherein the pH of the fermentation system during the fermentation in step 3) is between 3.5 and 6.0.

Description

Pichia kluyveri, pediococcus acidilactici and application of co-fermentation of pichia kluyveri and pediococcus acidilactici in degumming of fresh coffee beans Technical Field The invention belongs to the technical field of microbial fermentation, and particularly relates to a Kluyveromyces and Pediococcus acidilactici co-fermentation technology, in particular to Kluyveromyces, pediococcus acidilactici and application of co-fermentation in degumming fresh coffee beans. Background The main processing modes of coffee include a dry method, a semi-dry method and a wet method. In these treatments, fermentation is a key element, the main purpose of which is to remove the mucilage adhering to the parchment after the coffee has been peeled. In coffee fermentation, the combined fermentation of microorganisms has been attracting attention in recent years, and lactic acid bacteria and yeasts are widely used in the field of food industry in fermentation processes of foods such as grains, fruits and vegetables, bean products, dairy products, and the like. Besides the fast growth and reproduction and easy expansion culture, the microorganism has the advantages of accelerating the degradation of coffee mucilage to form good flavor and the like, and replaces natural fermentation degumming, chemical degumming, physical degumming and enzymatic degumming. The wet fermentation degumming mode adopted by the Yunnan small-grain coffee is easy to cause the problems of low coffee degumming efficiency, unstable coffee bean quality and environmental pollution caused by wastewater discharge, and meanwhile, the microbial pectase has great application potential in industries such as food, textile industry, paper industry and the like, so the microbial pectase is particularly important to the excavation of pectase-producing microorganisms. The microbial strain capable of producing the pectase at high yield is excavated, and the microbial strain is applied to the coffee fermentation process, so that the degumming effect and the influence of the degumming effect on coffee flavor substances are clear, and the microbial strain has important significance in developing a novel microbial degumming process for small-grain coffee, improving the quality of coffee beans, reducing environmental pollution and improving the flavor quality of the coffee beans. Disclosure of Invention The invention aims to provide a Kluyveromyces lactis and a pichia lactis and the application of co-fermentation thereof in degumming fresh coffee beans, and based on the purposes, the invention screens out two strains of high-yield pectase of the pichia lactis and the pichia kluyveromyces from different coffee processing modes, the co-fermentation technology of the pediococcus acidilactici and the pichia kluyveri is used for promoting the cooperation or influence of the pediococcus acidilactici and the pichia kluyveri, so that different effects on the degradation process of coffee mucilage and the flavor and quality of the final coffee are generated from single strain fermentation. The method comprises the following steps: First, the first object of the present invention is to obtain a strain of Pichia kluyveri (LII 35). A second object of the invention is to obtain a strain of Pediococcus acidilactici (KI 23). The third object of the invention is to provide the application of the pediococcus acidilactici KI23 and the Pichia kluyveri LII35 strain in biological degumming of coffee. The fourth object of the invention is to provide an application of the KI23 strain and the LII35 strain in preparing a preparation for biological degumming of coffee, wherein the preparation is a coffee bean degumming agent, the coffee bean degumming agent is mixed seed bacterial suspension of Pichia pastoris (Pichia kluyveri) with a strain number of LII35 and Pediococcus acidilactici (Pediococcus acidilactici) with a strain number of KI23, the Pichia pastoris LII35 is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No. 35115 and a preservation address of North Star Xiya No. 1 in the Beijing area, the Pediococcus acidilactici KI23 is preserved in China general microbiological culture collection center with a preservation number of CGMCC No. 35114 and a preservation address of North Star Xiya No. 1 in the Beijing area in the 2025 and 7 days. Further, the compounding ratio of the Kluyveromyces and the pediococcus acidilactici in the mixed seed bacterial suspension is (3-1) (1-3). Further, the bacterial liquid concentration of the mixed seed bacterial suspension is 10 5-109 CFU/mL. A fifth object of the invention is to provide a method for biological degumming of coffee using the KI23 strain and LII35 strain. The above object of the present invention is achieved by the following technical scheme: The invention separates a Kluyveri Pichia pastoris (Pichia kluyveri) LII35 strain from a honey-treated tim coffee (Coffea arabica ×C. canephora) samp