WO2021077788A1 - 一株解脂耶式酵母及其在制备低糖低脂椰蓉营养粉中的用途 - Google Patents

一株解脂耶式酵母及其在制备低糖低脂椰蓉营养粉中的用途 Download PDF

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WO2021077788A1
WO2021077788A1 PCT/CN2020/098116 CN2020098116W WO2021077788A1 WO 2021077788 A1 WO2021077788 A1 WO 2021077788A1 CN 2020098116 W CN2020098116 W CN 2020098116W WO 2021077788 A1 WO2021077788 A1 WO 2021077788A1
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coconut
low
fermentation
sugar
fat
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PCT/CN2020/098116
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French (fr)
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李静
邓毛程
李胜
王瑶
吴丰裕
蔡亮
顾继东
黄怀兴
吴林杰
王富程
李嘉俊
吴志东
黄洁华
廖民聪
林铿淳
邹小娜
赵祥源
谢杰文
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广东轻工职业技术学院
李静
邓毛程
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Priority to US17/770,631 priority Critical patent/US20220287342A1/en
Publication of WO2021077788A1 publication Critical patent/WO2021077788A1/zh

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
    • A23L25/40—Fermented products; Products treated with microorganisms or enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/165—Yeast isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
    • A23L25/30—Mashed or comminuted products, e.g. pulp, pastes, meal, powders; Products made therefrom, e.g. blocks, flakes, snacks; Liquid or semi-liquid products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

Definitions

  • the invention belongs to the technical field of microorganisms and food processing, and in particular relates to a Yarrowia lipolytica strain and its use in preparing low-sugar and low-fat coconut nutrient powder.
  • coconut belongs to the genus Palmaceae and is one of the main woody oil crops in tropical regions.
  • the world's coconut producing areas are Indonesia, the Philippines, India, Sri Lanka, Malaysia, Thailand and my country's Hainan province, with a total output of about 6 billion.
  • Coconut paste is the remaining part of the coconut meat after squeezing the juice. Under normal circumstances, the composition of coconut paste is about 3% moisture, about 45% oil, about 2% sugar, about 8% protein, about 40% dietary fiber, and about 2% ash. .
  • coconut is rich in dietary fiber and protein, and has extremely high nutritional value and physiological functions. With the improvement of people's living standards, excessive diet and excessive intake of high-energy substances have resulted in many "sub-health" and "riches disease” phenomena. Dietary fiber has a good promoting effect on improving the current dietary structure, and has received widespread attention, and is known as the seventh nutrient of civilization. A large number of studies have shown that dietary fiber has the functions of improving colon peristalsis, preventing constipation, removing intestinal toxins, preventing colon cancer, lowering blood pressure, lowering blood sugar, and lowering cholesterol. Coconut protein contains 18 kinds of amino acids, the ratio of essential amino acids is reasonable, and L-arginine is as high as 14.8%. researchers have found that coconut protein has the effects of lowering blood fat, lowering cholesterol, and improving immunity.
  • coconut paste is mainly used as fillings for baked goods, and the price is relatively low.
  • it is necessary to make full use of the dietary fiber and protein in coconut coconut to develop high-value nutritious food. Due to the rich oil content of coconut paste, it is not suitable for direct use as a nutritious food, and coconut oil needs to be removed.
  • the inventors of the present invention found a Yeast Yeast that has excellent ability to decompose coconut oil in the research on the treatment of coconut oil. Therefore, the primary objective of the present invention is to provide a Yarrowia lipolytica strain.
  • Another object of the present invention is to provide the use of the said Yarrowia lipolytica in the food field.
  • Another object of the present invention is to provide a low-sugar and low-fat coconut nutrient powder.
  • Yarrowia lipolytica YD19 A strain of Yarrowia lipolytica, named Yarrowia lipolytica YD19, and the deposit number is GDMCC No:60782. It was deposited on September 20, 2019 in Building 59, No. 100, Xianlie Middle Road, Guangzhou City. Lou Guangdong Microbial Culture Collection, Guangdong Institute of Microbiology.
  • the said lipolytic yeast can use sugar and fat as carbon sources, has excellent fat decomposition performance, and is suitable for application in the food field; and is more suitable for preparing low-sugar and low-fat coconut nutrient powder.
  • the fat is preferably coconut oil.
  • a low-sugar and low-fat coconut nutrient powder is prepared by the following steps:
  • Pretreatment of the coconut paste Mix the coconut paste with water and heat to soften the paste; then the heat-treated coconut paste is beaten, the obtained coconut paste is homogenized, the pH value is adjusted, and the paste is sterilized to obtain a suspension of coconut paste. liquid;
  • the water described in step (1) is preferably purified water.
  • the heating conditions described in step (1) are preferably: heating at 120 to 130°C for 30 to 60 minutes.
  • the conditions for beating in step (1) are preferably: homogenization at a rotation speed of 6000-8000 r/min.
  • the pressure of the homogenization in step (1) is preferably 30-40 MPa.
  • the number of times of the homogenization is preferably 2 to 3 times.
  • the pH value is preferably 6.0 to 7.0.
  • the sterilization conditions described in step (1) are preferably: sterilization at 115-121°C for 10-30 minutes; more preferably at 121°C for 10-15 minutes.
  • the Y. lipolytica culture solution described in step (2) is preferably a shake flask culture solution; it is preferably prepared by the following steps: inoculate Y. lipolytica in a shake flask culture medium, shake culture, and obtain shake flask culture liquid.
  • the shake flask culture medium is preferably formulated with glucose, yeast extract powder, malt extract powder, KH 2 PO 4 and MgSO 4 ⁇ 7H 2 O, etc.
  • the formula is preferably: glucose 25 ⁇ 35g/L, yeast extract powder 3 ⁇ 5g/L, malt extract powder 4-6g/L, KH 2 PO 4 1-3g/L, MgSO 4 ⁇ 7H 2 O 0.3-0.5g/L, adjust pH 5.5-6.5; more preferably: glucose 30g/ L, yeast extract powder 4g/L, malt extract powder 5g/L, KH 2 PO 4 2g/L, MgSO 4 ⁇ 7H 2 O 0.4g/L, adjust pH 6.0.
  • the amount of said shake flask culture medium in the shake flask is preferably 9%-20% of the volume of the shake flask; more preferably 10%-20%.
  • the conditions of the shaking culture are preferably: culture at 26-28°C and 160-200 r/min for 18-20 hours; more preferably: culture at 27-28°C and 160-200 r/min for 18-20 hours.
  • the inoculum of the culture solution of Y. lipolytica in step (2) is preferably 2%-10% (v/v).
  • the fermentation conditions described in step (2) are preferably: the stirring speed is 140-200r/min, the fermentation temperature is controlled to be 26-28°C, the aeration ratio of the fermentation is 0.16-0.48vvm, and the fermentation time is 96-120h; It is preferably as follows: the stirring speed is 140-200r/min, the fermentation temperature is controlled at 26-28°C, the fermentation time is 96-120h, and the aeration ratio of fermentation is gradually increased from 0.16vvm to 0.48vvm during the 0-36h of the fermentation period, In the last 24 hours of fermentation, the aeration ratio of the fermentation was controlled to 0.32 vvm, and the aeration ratio of the intermediate fermentation was controlled to 0.48 vvm.
  • the sterilization conditions described in step (2) are preferably: sterilization at 105-110°C for 10-15 minutes.
  • the conditions for the vacuum evaporation and concentration described in step (2) are preferably: the degree of vacuum is 0.07-0.08 MPa.
  • the volume after concentration by evaporation under reduced pressure is 30%-40% of the original volume.
  • step (2) The spray drying conditions described in step (2) are preferably: the inlet air temperature is controlled to be 160-200°C, and the outlet air temperature is controlled to be 70-90°C.
  • the degree of pulverization in step (2) is preferably pulverization until the particles reach 200-300 mesh.
  • the Yarrowia lipolytica provided by the present invention can make full use of the oil and sugar in the coconut paste as a carbon source.
  • Yeast is rich in protein. Its protein contains 8 essential amino acids, and the ratio of amino acids is close to the high-quality protein standard recommended by the Food and Agriculture Organization of the United Nations (FAO).
  • yeast also contains the complete B vitamins required by the human body, as well as rich ⁇ -1,3-glucan, as well as various minerals such as zinc, iron, magnesium, copper, selenium, and chromium.
  • the yeast is further processed and utilized on the coconut paste, and the obtained product is rich in yeast, coconut dietary fiber and coconut protein, which can improve the economic value of the yeast and coconut paste.
  • the present invention utilizes lipolytic yeast to degrade the oil and sugar in the coconut paste to prepare a fine coconut paste nutrient powder, which is rich in nutrition, low in sugar and fat, and has a strong coconut flavor, and is suitable as a prepared food for all people.
  • Figure 1 is a scanning electron micrograph of the bacterial cell of the present invention.
  • Figure 2 is a process flow diagram of the present invention.
  • the present invention will be further described in detail below in conjunction with the examples and drawings, but the implementation of the present invention is not limited thereto.
  • the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.
  • the reagents and materials used in the present invention are all commercially available.
  • Acclimation medium coconut oil 50g/L, yeast extract 10g/L, KH 2 PO 4 2.5g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, adjusted to pH 6.0.
  • the acclimation medium was sterilized at 121°C for 20 minutes. After cooling, the inoculum was 10% by volume, connected to the naturally fermented coconut milk, and cultured at 28°C and 200 rpm for 96 hours. Then, according to the inoculum volume of 10% by volume, the acclimation culture solution was transferred to fresh acclimation culture medium, and the culture was cultured for another 96 hours.
  • Plate screening medium peptone 20g/L, beef extract 10g/L, chloramphenicol 100mg/L, agar 20g/L, adjusted to pH 6.0.
  • the plate screening medium was sterilized at 121°C for 20 minutes, poured into a sterile petri dish while it was hot, and condensed to form a plate. Under aseptic operating conditions, spread sterile coconut oil on the plate, and then spread the gradient dilution of the acclimatization culture solution on the plate, and incubate at 28°C for 72 hours.
  • the slant medium adopts YEPD medium. Under aseptic operating conditions, use an inoculation needle to pick 60 single colonies on the plate, connect them to the slant medium, and cultivate them at 28°C. After the lawn grows, store them in the refrigerator at 4°C for later use.
  • Shake flask screening medium coconut oil 50g/L, yeast extract 10g/L, KH 2 PO 4 2.5g/L, MgSO 4 ⁇ 7H 2 O 0.5g/L, adjust pH 6.0.
  • the shake flask culture medium was sterilized at 121°C for 20 minutes. After cooling, 60 strains of slant strains were inserted into each flask, and each flask was inserted into a loop, and then cultured at 28°C and 200 rpm for 120 hours. Centrifuge the shake flask culture solution at 6000 r/min for 20 min, and collect the supernatant.
  • the residual oil in the supernatant was extracted twice with an equal volume of n-hexane, the extract layers were combined, and the n-hexane in the extract layer was removed by a rotary vacuum evaporator at 60°C, the mass of the residual oil was measured, and the degradation rate of coconut oil was calculated , The strain with the highest degradation rate is screened out, which is the strain of the present invention.
  • the strain was named Yarrowia lipolytica YD19, and it was preserved on September 20, 2019 at the Guangdong Provincial Institute of Microbiology, 5th Floor, Building 59, Compound, No. 100 Xianlie Middle Road, Guangzhou
  • the preservation center, the preservation number is GDMCC No:60782.
  • each bottle contains 150 mL of liquid, sterilized at 121°C for 20 minutes. After cooling, insert 1 ring of Yarrowia lipolytica GDMCC No:60782 slant bacterial lawn, and place it at 180r/min and 27°C. After culturing for 19 hours, the yeast shake flask culture solution is obtained.
  • the 1.8L yeast shake flask culture solution was connected to the fermentor, stirring was started and sterile air was introduced, and the fermentation was carried out for 96 hours.
  • the stirring speed of the fermenter is controlled to 170r/min, and the fermentation temperature is controlled to 27-28°C; according to the fermentation demand for dissolved oxygen, the aeration ratio of fermentation 0-24h is gradually increased from 0.16vvm to 0.48vvm, and the fermentation is 24 hours.
  • the aeration ratio of ⁇ 72h was controlled to 0.48vvm, and the aeration ratio after 72h of fermentation was controlled to 0.32vvm.
  • the fermentation broth was sterilized at 105° C. for 15 minutes, and then the volume of the fermentation broth was evaporated and concentrated to 30% volume under the condition of a vacuum of 0.07 MPa, and then spray-dried.
  • the inlet air temperature is controlled to be 190-200°C and the outlet air temperature is 80-90°C to obtain 542 g of a dried product with a moisture content of 2.2%.
  • the protein content of the coconut nutrient powder is 21.8%
  • the dietary fiber content is 59.4%
  • the content of B vitamins is 0.016%
  • fat content was 0.53%
  • glucose, fructose, sucrose, maltose, etc. were not detected.
  • Low-sugar and low-fat coconut nutrient powder products are prepared with water and have a strong coconut flavor.
  • each bottle contains 200mL of liquid, sterilized at 121°C for 20min. After cooling, insert 1 ring of Yarrowia lipolytica GDMCC No:60782 slant bacterial lawn, and place it at 200r/min and 28°C. After culturing for 18 hours, the yeast shake flask culture solution is obtained.
  • the fermentation broth was sterilized at 110° C. for 10 minutes, and then the volume of the fermentation broth was evaporated and concentrated to 40% volume under the condition of a vacuum of 0.075 MPa, and then spray-dried.
  • the inlet air temperature is controlled to be 170-180°C and the outlet air temperature is 75-85°C to obtain 676 g of a dried product with a moisture content of 2.4%.
  • the protein content of the coconut nutrient powder is 24.4%
  • the dietary fiber content is 57.4%
  • the content of B vitamins is 0.017%
  • fat content was 0.54%
  • glucose, fructose, sucrose, maltose, etc. were not detected.
  • Low-sugar and low-fat coconut nutrient powder products are prepared with water and have a strong coconut flavor.
  • each bottle contains 100 mL of liquid, sterilized at 121°C for 20 minutes. After cooling, insert 1 ring of Yarrowia lipolytica GDMCC No:60782 slant bacterial lawn, and place it at 200r/min and 28°C. After culturing for 20 hours, the yeast shake flask culture solution is obtained.
  • the stirring speed of the fermentation tank is controlled to 200r/min, and the fermentation temperature is controlled to 27-28°C; according to the fermentation demand for dissolved oxygen, the aeration ratio of fermentation 0 ⁇ 36h is gradually increased from 0.16vvm to 0.48vvm, and the fermentation is 36
  • the aeration ratio of ⁇ 96h was controlled to 0.48vvm, and the aeration ratio after 96h fermentation was controlled to 0.32vvm.
  • the fermentation broth was sterilized at 108° C. for 12 minutes, and then the volume of the fermentation broth was evaporated and concentrated to 35% volume under the condition of a vacuum of 0.08 MPa, and then spray-dried.
  • the inlet air temperature is controlled to 160-170°C and the outlet air temperature is 70-80°C to obtain 637 g of a dried product with a moisture content of 2.7%.
  • the protein content of the coconut nutrient powder is 23.6%
  • the dietary fiber content is 58.1%
  • the content of B vitamins is 0.017%
  • fat content was 0.53%
  • glucose, fructose, sucrose, maltose, etc. were not detected.
  • Low-sugar and low-fat coconut nutrient powder products are prepared with water and have a strong coconut flavor.
  • the coconut paste suspension is not fermented, it is directly evaporated and concentrated to 30% by volume under the condition of a vacuum degree of 0.07 MPa, and then spray-dried using a spray drying tower.
  • the inlet air temperature is controlled to be 190-200°C and the outlet air temperature is 80-90°C. Due to the high oil content, dry powder cannot be obtained.
  • each bottle contains 150 mL of liquid, sterilized at 121°C for 20 minutes. After cooling, insert 1 ring of Saccharomyces cerevisiae CICC1252 (purchased from China Industrial Microbial Culture Collection and Management Center). Cultivate for 19 hours under the conditions of 180r/min and 27°C to obtain yeast shake flask culture solution.
  • the 1.8L yeast shake flask culture solution was connected to the fermentor, stirring was started and sterile air was introduced, and the fermentation was carried out for 96 hours.
  • the stirring speed of the fermenter is controlled to 170r/min, and the fermentation temperature is controlled to 27-28°C; according to the fermentation demand for dissolved oxygen, the aeration ratio of fermentation 0-24h is gradually increased from 0.16vvm to 0.48vvm, and the fermentation is 24 hours.
  • the aeration ratio of ⁇ 72h was controlled to 0.48vvm, and the aeration ratio after 72h of fermentation was controlled to 0.32vvm.
  • the fermentation broth was sterilized at 105° C. for 15 minutes, and then the volume of the fermentation broth was evaporated and concentrated to 30% volume under the condition of a vacuum of 0.07 MPa, and then spray-dried.
  • the inlet air temperature is controlled to be 190-200°C and the outlet air temperature is 80-90°C. Due to the high oil content, dry powder cannot be obtained.
  • each bottle contains 150 mL of liquid, sterilized at 121°C for 20 minutes, and after cooling, insert 1 ring of Yarrowia lipolytica CICC31120 (purchased from the China Industrial Microbial Culture Collection and Management Center). The moss was cultured for 19 hours under the conditions of 180r/min and 27°C to obtain the yeast shake flask culture solution.
  • the 1.8L yeast shake flask culture solution was connected to the fermentor, stirring was started and sterile air was introduced, and the fermentation was carried out for 96 hours.
  • the stirring speed of the fermenter is controlled to 170r/min, and the fermentation temperature is controlled to 27-28°C; according to the fermentation demand for dissolved oxygen, the aeration ratio of fermentation 0-24h is gradually increased from 0.16vvm to 0.48vvm, and the fermentation is 24 hours.
  • the aeration ratio of ⁇ 72h was controlled to 0.48 vvm, and the aeration ratio after 72h of fermentation was controlled to 0.32 vvm.
  • the fermentation broth was sterilized at 105° C. for 15 minutes, and then the volume of the fermentation broth was evaporated and concentrated to 30% volume under the condition of a vacuum of 0.07 MPa, and then spray-dried.
  • the inlet air temperature is controlled to be 190-200°C and the outlet air temperature is 80-90°C. Due to the high oil content, dry powder cannot be obtained.
  • each bottle contains 150 mL of liquid, sterilized at 121°C for 20 minutes. After cooling, connect 1 ring of Candida utilis CICC31126 (purchased from the China Industrial Microbial Culture Collection and Management Center). The moss was cultured for 19 hours under the conditions of 180r/min and 27°C to obtain the yeast shake flask culture solution.
  • the 1.8L yeast shake flask culture solution was connected to the fermentor, stirring was started and sterile air was introduced, and the fermentation was carried out for 96 hours.
  • the stirring speed of the fermenter is controlled to 170r/min, and the fermentation temperature is controlled to 27-28°C; according to the fermentation demand for dissolved oxygen, the aeration ratio of fermentation 0-24h is gradually increased from 0.16vvm to 0.48vvm, and the fermentation is 24 hours.
  • the aeration ratio of ⁇ 72h was controlled to 0.48vvm, and the aeration ratio after 72h of fermentation was controlled to 0.32vvm.
  • the inlet air temperature is controlled to be 190-200°C and the outlet air temperature is 80-90°C. Due to the high oil content, dry powder cannot be obtained.
  • each bottle contains 150 mL of liquid, sterilized at 121°C for 20 minutes, and after cooling, insert 1 ring of Yarrowia lipolytica CICC32520 (purchased from the China Industrial Microbial Culture Collection and Management Center). The moss was cultured for 19 hours under the conditions of 180r/min and 27°C to obtain the yeast shake flask culture solution.
  • the 1.8L yeast shake flask culture solution was connected to the fermentor, stirring was started and sterile air was introduced, and the fermentation was carried out for 96 hours.
  • the stirring speed of the fermenter is controlled to 170r/min, and the fermentation temperature is controlled to 27-28°C; according to the fermentation demand for dissolved oxygen, the aeration ratio of fermentation 0-24h is gradually increased from 0.16vvm to 0.48vvm, and the fermentation is 24 hours.
  • the aeration ratio of ⁇ 72h was controlled to 0.48vvm, and the aeration ratio after 72h of fermentation was controlled to 0.32vvm.
  • the inlet air temperature is controlled to be 190-200°C and the outlet air temperature is 80-90°C. Due to the high oil content, dry powder cannot be obtained.

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Abstract

提供了一株解脂耶式酵母及其在制备低糖低脂椰蓉营养粉中的用途，该解脂耶式酵母的保藏编号为GDMCC No:60782。通过将椰蓉与水混合，加热；接着将加热处理后的椰蓉打浆，将得到的椰浆均质，调节pH值，灭菌，得到椰蓉悬液；在冷却的椰蓉悬液中接入所述解脂耶式酵母的培养液，发酵；将得到的发酵液灭菌，喷雾干燥，粉碎，得到低糖低脂椰蓉营养粉。

Description

一株解脂耶式酵母及其在制备低糖低脂椰蓉营养粉中的用途 技术领域
本发明属于微生物和食品加工技术领域，特别涉及一株解脂耶式酵母及其在制备低糖低脂椰蓉营养粉中的用途。
背景技术
椰子属于棕榈科椰子属，是热带地区主要的木本油料作物之一。全世界椰子产区是印度尼西亚、菲律宾、印度、斯里兰卡、马来西亚、泰国和我国的海南省，总产量约为60亿个。椰蓉是椰肉榨汁后的剩余部分，一般情况下，椰蓉的成分为：水分3％左右，油脂45％左右，糖分2％左右，蛋白质8％左右，膳食纤维40％左右，灰分2％左右。
椰蓉含有丰富的膳食纤维和蛋白质，具有极高的营养价值和生理功能。随着人们生活水平的提高，过度饮食和高能量物质过多摄入，产生很多“亚健康”和“富贵病”现象。膳食纤维对改善当前膳食结构有较好的促进作用，受到了广泛关注，被誉为人类第七种营养素。大量研究表明，膳食纤维具有改善大肠蠕动、防止便秘、清除肠道毒素、预防结肠癌、降低血压、降低血糖、降低胆固醇等功能。椰子蛋白含有18种氨基酸，必需氨基酸配比合理，L-精氨酸高达14.8％，研究人员发现椰子蛋白具有降低血脂、降低胆固醇、提高免疫力等功效。
目前，椰蓉主要用于烘焙食品的馅料，价格比较低。为了开拓椰蓉的高附加值利用途径，有必要充分利用椰蓉中的膳食纤维和蛋白质，开发高价值的营养食品。由于椰蓉的油脂含量丰富，不适宜直接作为营养食品，需对椰子油脂进行去除处理。
目前，尚未生物方法对椰蓉油脂处理的相关报道。
发明内容
为了创新椰蓉营养粉产品，解决椰蓉的低值应用问题，本发明发明人在对椰蓉油脂处理的研究中，发现一株解脂耶式酵母，其分解椰子油的能力优异。从而，本发明的首要目的在于提供一株解脂耶式酵母。
本发明的另一目的在于提供所述的解脂耶式酵母在食品领域中的用途。
本发明的再一目的在于提供一种低糖低脂椰蓉营养粉。
为实现上述目的，本发明通过下述技术方案实现：
一株解脂耶式酵母，名称为解脂耶式酵母(Yarrowia lipolytica)YD19，保藏编号为GDMCC No:60782，于2019年9月20日保藏于位于广州市先烈中路100号大院59栋5楼广东省微生物研究所的广东省微生物菌种保藏中心。
所述的解脂耶式酵母能以糖和脂肪作为碳源，分解脂肪的性能优异，适合在食品领域中进行应用；更适合用于制备低糖低脂椰蓉营养粉。
所述的脂肪优选为椰子油。
一种低糖低脂椰蓉营养粉，通过如下步骤制备得到：
(1)椰蓉的预处理：将椰蓉与水混合，加热，目的是为了使椰蓉软化；接着将加热处理后的椰蓉打浆，将得到的椰浆均质，调节pH值，灭菌，得到椰蓉悬液；
(2)发酵与后处理：将上述解脂耶式酵母的培养液接种于冷却的椰蓉悬液中，发酵；将得到的发酵液灭菌，减压蒸发浓缩，喷雾干燥，粉碎，得到低糖低脂椰蓉营养粉。
步骤(1)中所述的水优选为纯净水。
所述的水的用量优选按椰蓉：水＝质量比1：(22～24)配比计算。
步骤(1)中所述的加热的条件优选为：于120～130℃加热30～60min。
步骤(1)中所述的打浆的条件优选为：于6000～8000r/min的转速匀浆。
步骤(1)中所述的均质的压力优选为30～40MPa。
所述的均质的次数优选为2～3次。
所述的pH值优选为6.0～7.0。
步骤(1)中所述的灭菌的条件优选为：于115～121℃灭菌10～30min；更优选为于121℃灭菌10～15min。
步骤(2)中所述的解脂耶式酵母培养液优选为摇瓶培养液；优选通过如下步骤制备得到：将解脂耶式酵母接种于摇瓶培养基中，振荡培养，得到摇瓶培养液。
所述的摇瓶培养基优选葡萄糖、酵母浸粉、麦芽浸粉、KH 2PO 4和MgSO 4·7H 2O等配制而成，配方优选为：葡萄糖25～35g/L，酵母浸粉3～5g/L，麦芽浸粉4～6g/L，KH 2PO 4 1～3g/L，MgSO 4·7H 2O 0.3～0.5g/L，调节pH5.5～6.5；更优选为：葡萄糖30g/L，酵母浸粉4g/L，麦芽浸粉5g/L，KH 2PO 4 2g/L，MgSO 4·7H 2O 0.4g/L，调节pH6.0。
所述的摇瓶培养基在摇瓶中的量优选为摇瓶容积的9％～20％；更优选为10％～20％。
所述的振荡培养的条件优选为：于26～28℃、160～200r/min培养18～20h；更优选为： 于27～28℃、160～200r/min培养18～20h。
步骤(2)中所述的解脂耶式酵母的培养液的接种量优选为2％～10％(v/v)。
步骤(2)中所述的发酵的条件优选为：搅拌转速为140～200r/min，发酵温度控制为26～28℃，发酵的通气比为0.16～0.48vvm，发酵时间为96～120h；更优选如下：搅拌转速为140～200r/min，发酵温度控制为26～28℃，发酵时间为96～120h，在发酵期间的第0～36h，发酵的通气比由0.16vvm逐步增加至0.48vvm，发酵最后的24h，发酵的通气比控制为0.32vvm，中间的发酵的通气比控制为0.48vvm。
步骤(2)中所述的灭菌的条件优选为：于105～110℃灭菌10～15min。
步骤(2)中所述的减压蒸发浓缩的条件优选为：真空度为0.07～0.08MPa。
所述的减压蒸发浓缩后的体积为原体积的30％～40％。
步骤(2)中所述的喷雾干燥条件优选为：进风温度控制为160～200℃，出风温度控制为70～90℃。
步骤(2)中所述的粉碎的程度优选为粉碎至微粒达到200～300目。
本发明具有以下优点及效果：
(1)本发明提供的解脂耶式酵母能充分利用椰蓉中的油脂和糖分作为碳源。酵母含有丰富的蛋白质，其蛋白质含有人体必需的8种氨基酸，且氨基酸配比接近联合国粮农组织(FAO)推荐的优质蛋白标准。另外，酵母还含有人体所需的完整B族维生素，以及丰富的β-1,3-葡聚糖，以及锌、铁、镁、铜、硒、铬等多种矿物质。将该酵母对椰蓉进一步加工利用，得到的产品中富含酵母、椰子膳食纤维和椰子蛋白，时提升酵母和椰蓉的经济价值。
(2)由于椰蓉中油脂含量较高，无法将其制成微细的干粉。本发明利用解脂耶式酵母降解椰蓉中的油脂和糖分，制备得到微细的椰蓉营养粉，营养丰富，低糖低脂，椰香风味浓郁，适宜作为所有人群的冲调食品。
附图说明
图1是本发明的菌体扫描电镜照片图。
图2是本发明的工艺流程图。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述，但本发明的实施方式不限于此。除 非特别说明，本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。除非特别说明，本发明所用试剂和材料均可通过市售获得。
实施例1
以自然发酵5天的椰浆为菌种筛选材料。
驯化培养基：椰子油50g/L，酵母膏10g/L，KH 2PO 4 2.5g/L，MgSO 4·7H 2O 0.5g/L，调节pH6.0。驯化培养基于121℃灭菌20min，冷却后，按体积比10％的接种量，接入自然发酵的椰浆，于28℃、200rpm的条件下培养96h。然后，再按体积比10％的接种量，将驯化培养液转接入新鲜的驯化培养基，再培养96h。
平板筛选培养基：蛋白胨20g/L，牛肉膏10g/L，氯霉素100mg/L，琼脂20g/L，调节pH6.0。平板筛选培养基于121℃灭菌20min，趁热倒入无菌的培养皿，冷凝形成平板。在无菌操作条件下，将无菌的椰子油涂布在平板上，再将驯化培养液的梯度稀释液涂布于平板上，置于28℃下培养72h。
斜面培养基采用YEPD培养基。在无菌操作条件下，用接种针挑取平板上60个单菌落，分别接入斜面培养基，于28℃下培养，菌苔长出后，置于4℃冰箱保藏，备用。
摇瓶筛选培养基：椰子油50g/L，酵母膏10g/L，KH 2PO 4 2.5g/L，MgSO 4·7H 2O 0.5g/L，调节pH6.0。摇瓶培养基于121℃灭菌20min，冷却后，分别接入60株斜面菌种，每瓶接入1环，然后置于28℃、200rpm的条件下培养120h。将摇瓶培养液于6000r/min的条件下离心20min，收集上清液。用等体积的正己烷对上清液中残余油脂进行萃取两次，合并萃取层，利用旋转真空蒸发器于60℃去除萃取层的正己烷，测定残余油脂的质量，计算出椰子油的降解率，筛选出降解率最高的一株菌种，即为本发明的菌种。
利用扫描电镜观察该菌种的菌体形态，菌体形态与普通酵母的菌体形态一致(如图1所示)。利用分子鉴定方法分析该菌种的26S rDNA，在NCBI数据平台上检索与比对，确定该菌种为解脂耶式酵母(Yarrowia lipolytica)。该菌种的26S rDNA如下所示。
Figure PCTCN2020098116-appb-000001
Figure PCTCN2020098116-appb-000002
将该菌种命名为解脂耶式酵母(Yarrowia lipolytica)YD19，于2019年9月20日保藏于位于广州市先烈中路100号大院59栋5楼广东省微生物研究所的广东省微生物菌种保藏中心，保藏编号为GDMCC No:60782。
实施例2
一种低糖低脂椰蓉营养粉的制作过程，如图2所示，具体如下：
(1)椰蓉的预处理
称取0.75kg干椰蓉，加入18kg纯净水，置于130℃的条件下加热30min，使椰蓉软化。在6000r/min的条件下，利用高速匀浆机将混合液打成浆状，然后利用高压均质机在30MPa的压力下进行3次均质，调节pH至7.0，放入发酵罐内。在121℃下灭菌10min，冷却至室温，备用。
(2)椰蓉悬液的发酵与后处理
称取54g葡萄糖、7.2g酵母浸粉、9g麦芽浸粉、3.6g KH 2PO 4、0.72g MgSO 4·7H 2O，加水溶解，定容1.8L，调节pH至6.0，分装至12个1000mL三角瓶中，每瓶装液150mL，121℃灭菌20min，冷却后，接入1环解脂耶式酵母GDMCC No:60782斜面菌种的菌苔，置于180r/min、27℃的条件下进行培养19h，即得酵母摇瓶培养液。
将1.8L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵96h。在发酵过程中，发酵罐搅拌转速控制为170r/min，发酵温度控制为27～28℃；根据发酵对溶解氧的需求，发酵0～24h的通气比由0.16vvm逐步增加至0.48vvm，发酵24～72h的通气比控制为0.48vvm，发酵72h后的通气比控制为0.32vvm。
发酵结束后，在105℃下对发酵液进行灭菌15min，然后将发酵液体积于真空度为0.07MPa的条件下进行蒸发浓缩至30％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为190～200℃，出风温度为80～90℃，获得542g含水分2.2％的干燥物。利用超微粉碎机将干燥物粉碎至200～300目，即为低糖低脂椰蓉营养粉，经检测，该椰蓉营养粉的蛋白质含量为21.8％，膳食纤维含量为59.4％，B族维生素含量为0.016％，脂肪含量为0.53％，葡萄糖、果糖、蔗糖、麦芽糖等没有被检出。低糖低脂椰蓉营养粉产品用水冲调，椰香风味浓郁。
实施例3
一种低糖低脂椰蓉营养粉的制作过程，如图2所示，具体如下：
(1)椰蓉的预处理
称取0.954kg干椰蓉，加入21kg纯净水，置于120℃的条件下加热60min，使椰蓉软化。在8000r/min的条件下，利用高速匀浆机将混合液打成浆状，然后利用高压均质机在40MPa的压力下进行2次均质，调节pH至6.0，放入发酵罐内。在121℃下灭菌15min，冷却至室温，备用。
(2)椰蓉悬液的发酵与后处理
称取42g葡萄糖、5.6g酵母浸粉、7g麦芽浸粉、2.8g KH 2PO 4、0.56g MgSO 4·7H 2O，加水溶解，定容1.4L，调节pH至6.0，分装至7个1000mL三角瓶中，每瓶装液200mL，121℃灭菌20min，冷却后，接入1环解脂耶式酵母GDMCC No:60782斜面菌种的菌苔，置于200r/min、28℃的条件下进行培养18h，即得酵母摇瓶培养液。
将1.4L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵108h。在发酵过程中，发酵罐搅拌转速控制为140r/min，发酵温度控制为26～27℃；根据发酵对溶解氧的需求，发酵0～30h的通气比由0.16vvm逐步增加至0.48vvm，发酵30～84h的通气比控制为0.48vvm，发酵84h后的通气比控制为0.32vvm。
发酵结束后，在110℃下对发酵液进行灭菌10min，然后将发酵液体积于真空度为0.075MPa的条件下进行蒸发浓缩至40％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为170～180℃，出风温度为75～85℃，获得676g含水分2.4％的干燥物。利用超微粉碎机将干燥物粉碎至200～300目，即为低糖低脂椰蓉营养粉，经检测，该椰蓉营养粉的蛋白质含量为24.4％，膳食纤维含量为57.4％，B族维生素含量为0.017％，脂肪含量为0.54％，葡萄糖、果糖、蔗糖、麦芽糖等没有被检出。低糖低脂椰蓉营养粉产品用水冲调，椰香风味浓郁。
实施例4
一种低糖低脂椰蓉营养粉的制作过程，如图2所示，具体如下：
(1)椰蓉的预处理
称取0.869kg干椰蓉，加入20kg纯净水，置于125℃的条件下加热45min，使椰蓉软化。在7000r/min的条件下，利用高速匀浆机将混合液打成浆状，然后利用高压均质机在35MPa的压力下进行3次均质，调节pH至6.5，放入发酵罐内。在121℃下灭菌12min，冷却至室温，备用。
(2)椰蓉悬液的发酵与后处理
称取12g葡萄糖、1.6g酵母浸粉、2g麦芽浸粉、0.8g KH 2PO 4、0.16g MgSO 4·7H 2O，加水溶解，定容0.4L，调节pH至6.5，分装至4个1000mL三角瓶中，每瓶装液100mL，121℃灭菌20min，冷却后，接入1环解脂耶式酵母GDMCC No:60782斜面菌种的菌苔，置于200r/min、28℃的条件下进行培养20h，即得酵母摇瓶培养液。
将0.4L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵120h。在发酵过程中，发酵罐搅拌转速控制为200r/min，发酵温度控制为27～28℃；根据发酵对溶解氧的需求，发酵0～36h的通气比由0.16vvm逐步增加至0.48vvm，发酵36～96h的通气比控制为0.48vvm，发酵96h后的通气比控制为0.32vvm。
发酵结束后，在108℃下对发酵液进行灭菌12min，然后将发酵液体积于真空度为0.08MPa的条件下进行蒸发浓缩至35％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为160～170℃，出风温度为70～80℃，获得637g含水分2.7％的干燥物。利用超微粉碎机将干燥物粉碎至200～300目，即为低糖低脂椰蓉营养粉，经检测，该椰蓉营养粉的蛋白质含量为23.6％，膳食纤维含量为58.1％，B族维生素含量为0.017％，脂肪含量为0.53％，葡萄糖、果糖、蔗糖、麦芽糖等没有被检出。低糖低脂椰蓉营养粉产品用水冲调，椰香风味浓郁。
对比例1
(1)椰蓉的预处理
称取0.75kg干椰蓉，加入18kg纯净水，置于130℃的条件下加热30min，使椰蓉软化。在6000r/min的条件下，利用高速匀浆机将混合液打成浆状，然后利用高压均质机在30MPa的压力下进行3次均质，调节pH至7.0。在121℃下灭菌10min，冷却至室温，备用。
(2)椰蓉悬液的后处理
椰蓉悬液未经发酵，直接于真空度为0.07MPa的条件下进行蒸发浓缩至30％体积，再利用喷雾干燥塔进行喷雾干燥。在喷雾干燥过程中，控制进风温度为190～200℃，出风温度为80～90℃，由于油脂含量高，无法获得干燥的粉体。
对比例2
(1)椰蓉的预处理
称取0.75kg干椰蓉，加入18kg纯净水，置于130℃的条件下加热30min，使椰蓉软化。在6000r/min的条件下，利用高速匀浆机将混合液打成浆状，然后利用高压均质机在30MPa的压力下进行3次均质，调节pH至7.0，放入发酵罐内。在121℃下灭菌10min，冷却至室 温，备用。
(2)椰蓉悬液的发酵与后处理
称取54g葡萄糖、7.2g酵母浸粉、9g麦芽浸粉、3.6g KH 2PO 4、0.72g MgSO 4·7H 2O，加水溶解，定容1.8L，调节pH至6.0，分装至12个1000mL三角瓶中，每瓶装液150mL，121℃灭菌20min，冷却后，接入1环酿酒酵母(Saccharomyces cerevisiae)CICC1252(购买于中国工业微生物菌种保藏管理中心)斜面菌种的菌苔，置于180r/min、27℃的条件下进行培养19h，即得酵母摇瓶培养液。
将1.8L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵96h。在发酵过程中，发酵罐搅拌转速控制为170r/min，发酵温度控制为27～28℃；根据发酵对溶解氧的需求，发酵0～24h的通气比由0.16vvm逐步增加至0.48vvm，发酵24～72h的通气比控制为0.48vvm，发酵72h后的通气比控制为0.32vvm。
发酵结束后，在105℃下对发酵液进行灭菌15min，然后将发酵液体积于真空度为0.07MPa的条件下进行蒸发浓缩至30％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为190～200℃，出风温度为80～90℃，由于油脂含量高，无法获得干燥的粉体。
对比例3
(1)椰蓉的预处理
称取0.75kg干椰蓉，加入18kg纯净水，置于130℃的条件下加热30min，使椰蓉软化。在6000r/min的条件下，利用高速匀浆机将混合液打成浆状，然后利用高压均质机在30MPa的压力下进行3次均质，调节pH至7.0，放入发酵罐内。在121℃下灭菌10min，冷却至室温，备用。
(2)椰蓉悬液的发酵与后处理
称取54g葡萄糖、7.2g酵母浸粉、9g麦芽浸粉、3.6g KH 2PO 4、0.72g MgSO 4·7H 2O，加水溶解，定容1.8L，调节pH至6.0，分装至12个1000mL三角瓶中，每瓶装液150mL，121℃灭菌20min，冷却后，接入1环解脂亚罗酵母(Yarrowia lipolytica)CICC31120(购买于中国工业微生物菌种保藏管理中心)斜面菌种的菌苔，置于180r/min、27℃的条件下进行培养19h，即得酵母摇瓶培养液。
将1.8L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵96h。在发酵过程中，发酵罐搅拌转速控制为170r/min，发酵温度控制为27～28℃；根据发酵对溶解氧的需求，发酵0～24h的通气比由0.16vvm逐步增加至0.48vvm，发酵24～72h的通气比控制为0.48 vvm，发酵72h后的通气比控制为0.32vvm。
发酵结束后，在105℃下对发酵液进行灭菌15min，然后将发酵液体积于真空度为0.07MPa的条件下进行蒸发浓缩至30％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为190～200℃，出风温度为80～90℃，由于油脂含量高，无法获得干燥的粉体。
对比例4
(1)椰蓉的预处理
称取0.75kg干椰蓉，加入18kg纯净水，置于130℃的条件下加热30min，使椰蓉软化。在6000r/min的条件下，利用高速匀浆机将混合液打成浆状，调节pH至7.0，放入发酵罐内。在121℃下灭菌10min，冷却至室温，备用。
(2)椰蓉悬液的发酵与后处理
称取54g葡萄糖、7.2g酵母浸粉、9g麦芽浸粉、3.6g KH 2PO 4、0.72g MgSO 4·7H 2O，加水溶解，定容1.8L，调节pH至6.0，分装至12个1000mL三角瓶中，每瓶装液150mL，121℃灭菌20min，冷却后，接入1环产朊假丝酵母(Candida utilis)CICC31126(购买于中国工业微生物菌种保藏管理中心)斜面菌种的菌苔，置于180r/min、27℃的条件下进行培养19h，即得酵母摇瓶培养液。
将1.8L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵96h。在发酵过程中，发酵罐搅拌转速控制为170r/min，发酵温度控制为27～28℃；根据发酵对溶解氧的需求，发酵0～24h的通气比由0.16vvm逐步增加至0.48vvm，发酵24～72h的通气比控制为0.48vvm，发酵72h后的通气比控制为0.32vvm。
发酵结束后，利用高压均质机在30MPa的压力下对发酵液进行3次均质，在105℃下对发酵液进行灭菌15min，然后将发酵液体积于真空度为0.07MPa的条件下进行蒸发浓缩至30％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为190～200℃，出风温度为80～90℃，由于油脂含量高，无法获得干燥的粉体。
对比例5
(1)椰蓉的预处理
称取0.75kg干椰蓉，加入18kg纯净水，置于130℃的条件下加热30min，使椰蓉软化。在6000r/min的条件下，利用高速匀浆机将混合液打成浆状，调节pH至7.0，放入发酵罐内。在121℃下灭菌10min，冷却至室温，备用。
(2)椰蓉悬液的发酵与后处理
称取54g葡萄糖、7.2g酵母浸粉、9g麦芽浸粉、3.6g KH 2PO 4、0.72g MgSO 4·7H 2O，加水溶解，定容1.8L，调节pH至6.0，分装至12个1000mL三角瓶中，每瓶装液150mL，121℃灭菌20min，冷却后，接入1环解脂耶式酵母(Yarrowia lipolytica)CICC32520(购买于中国工业微生物菌种保藏管理中心)斜面菌种的菌苔，置于180r/min、27℃的条件下进行培养19h，即得酵母摇瓶培养液。
将1.8L酵母摇瓶培养液接入发酵罐，启动搅拌和通入无菌空气，发酵96h。在发酵过程中，发酵罐搅拌转速控制为170r/min，发酵温度控制为27～28℃；根据发酵对溶解氧的需求，发酵0～24h的通气比由0.16vvm逐步增加至0.48vvm，发酵24～72h的通气比控制为0.48vvm，发酵72h后的通气比控制为0.32vvm。
发酵结束后，利用高压均质机在30MPa的压力下对发酵液进行3次均质，在105℃下对发酵液进行灭菌15min，然后将发酵液体积于真空度为0.07MPa的条件下进行蒸发浓缩至30％体积，再进行喷雾干燥。在喷雾干燥过程中，控制进风温度为190～200℃，出风温度为80～90℃，由于油脂含量高，无法获得干燥的粉体。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。
Figure PCTCN2020098116-appb-000003
Figure PCTCN2020098116-appb-000004

Claims (10)

  1. 一株解脂耶式酵母，其特征在于：所述的解脂耶式酵母的名称为解脂耶式酵母(Yarrowia lipolytica)YD19，保藏编号为GDMCC No:60782，于2019年9月20日保藏于位于广州市先烈中路100号大院59栋5楼广东省微生物研究所的广东省微生物菌种保藏中心。
  2. 权利要求1所述的解脂耶式酵母在食品领域中的应用。
  3. 根据权利要求2所述的解脂耶式酵母在食品领域中的应用，其特征在于：所述的解脂耶式酵母用于制备低糖低脂椰蓉营养粉。
  4. 一种低糖低脂椰蓉营养粉，其特征在于通过如下步骤制备得到：
    (1)椰蓉的预处理：将椰蓉与水混合，加热；接着将加热处理后的椰蓉打浆，将得到的椰浆均质，调节pH值，灭菌，得到椰蓉悬液；
    (2)发酵与后处理：将权利要求1所述的解脂耶式酵母的培养液接种于冷却的椰蓉悬液中，发酵；将得到的发酵液灭菌，减压蒸发浓缩，喷雾干燥，粉碎，得到低糖低脂椰蓉营养粉。
  5. 根据权利要求4所述的低糖低脂椰蓉营养粉，其特征在于：
    步骤(1)中所述的pH值为6.0～7.0；
    步骤(1)中所述的加热的条件为：于120～130℃加热30～60min；
    步骤(1)中所述的打浆的条件为：于6000～8000r/min的转速匀浆；
    步骤(1)中所述的均质的压力为30～40MPa；
    步骤(2)中所述的解脂耶式酵母培养液为摇瓶培养液；
    步骤(2)中所述的解脂耶式酵母的培养液的接种量为2％～10％(v/v)；
    步骤(2)中所述的发酵的条件为：搅拌转速为140～200r/min，发酵温度控制为26～28℃，发酵的通气比为0.16～0.48vvm，发酵时间为96～120h；
    步骤(2)中所述的减压蒸发浓缩的条件为：真空度为0.07～0.08MPa；
    步骤(2)中所述的喷雾干燥条件为：进风温度控制为160～200℃，出风温度控制为70～90℃。
  6. 根据权利要求5所述的低糖低脂椰蓉营养粉，其特征在于：
    所述的均质的次数为2～3次；
    所述的减压蒸发浓缩后的体积为原体积的30％～40％。
  7. 根据权利要求5所述的低糖低脂椰蓉营养粉，其特征在于：
    步骤(2)中所述的解脂耶式酵母培养液通过如下步骤制备得到：将解脂耶式酵母接种于摇瓶培养基中，振荡培养，得到摇瓶培养液；
    所述的摇瓶培养基的组分为：葡萄糖25～35g/L，酵母浸粉3～5g/L，麦芽浸粉4～6g/L，KH 2PO 41～3g/L，MgSO 4·7H 2O 0.3～0.5g/L，调节pH5.5～6.5；
    所述的摇瓶培养基在摇瓶中的量为摇瓶容积的9％～20％
    所述的振荡培养的条件为：于26～28℃、160～200r/min培养18～20h。
  8. 根据权利要求4所述的低糖低脂椰蓉营养粉，其特征在于：
    步骤(1)中所述的水为纯净水；
    所述的水的用量按椰蓉：水＝质量比1：(22～24)配比计算。
  9. 根据权利要求4所述的低糖低脂椰蓉营养粉，其特征在于：
    步骤(1)中所述的灭菌的条件为：于115～121℃灭菌10～30min；
    步骤(2)中所述的灭菌的条件为：于105～110℃灭菌10～15min。
  10. 根据权利要求4所述的低糖低脂椰蓉营养粉，其特征在于：步骤(2)中所述的粉碎的程度为粉碎至微粒达到200～300目。
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