WO2021012870A1 - 一种低嘌呤豆浆粉及其制作方法 - Google Patents

一种低嘌呤豆浆粉及其制作方法 Download PDF

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WO2021012870A1
WO2021012870A1 PCT/CN2020/098087 CN2020098087W WO2021012870A1 WO 2021012870 A1 WO2021012870 A1 WO 2021012870A1 CN 2020098087 W CN2020098087 W CN 2020098087W WO 2021012870 A1 WO2021012870 A1 WO 2021012870A1
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fermentation
purine
milk powder
soybean milk
malt extract
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PCT/CN2020/098087
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English (en)
French (fr)
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邓毛程
李静
谢俊宏
栗瑞敏
王瑶
黄怀兴
蔡亮
王富程
黄洁华
谭宏业
赵祥源
林铿淳
廖民聪
邹小娜
李建忠
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广东轻工职业技术学院
广东创新科技职业学院
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Priority to AU2020316414A priority Critical patent/AU2020316414B2/en
Publication of WO2021012870A1 publication Critical patent/WO2021012870A1/zh

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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C1/00Concentration, evaporation or drying
    • A23C1/12Concentration by evaporation
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C1/00Concentration, evaporation or drying
    • A23C1/14Concentration, evaporation or drying combined with other treatment
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C11/00Milk substitutes, e.g. coffee whitener compositions
    • A23C11/02Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
    • A23C11/10Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
    • A23C11/103Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
    • A23C11/106Addition of, or treatment with, microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C3/00Preservation of milk or milk preparations
    • A23C3/02Preservation of milk or milk preparations by heating
    • A23C3/03Preservation of milk or milk preparations by heating the materials being loose unpacked
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C7/00Other dairy technology
    • A23C7/04Removing unwanted substances other than lactose or milk proteins from milk
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C2210/00Physical treatment of dairy products
    • A23C2210/15High pressure treatment
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C2210/00Physical treatment of dairy products
    • A23C2210/20Treatment using membranes, including sterile filtration
    • A23C2210/206Membrane filtration of a permeate obtained by ultrafiltration, nanofiltration or microfiltration

Definitions

  • the invention belongs to the technical field of food processing, and particularly relates to a low-purine soybean milk powder and a preparation method thereof.
  • Soy milk is a beverage made by soaking, refining, filtering, and boiling soybeans. It is rich in nutrients such as protein, fat, minerals and vitamins. It also contains health functional substances such as soybean isoflavones and lecithin. "Plant milk" reputation. Making soy milk into soy milk powder can meet consumers' needs for brewing drinks on different occasions. However, whether it is freshly ground soy milk or soy milk prepared with soy milk powder, the total purine content is very high. At present, the total purine content of most commercially available soybean milk powder (commodities without sugar) is 1800-2600 mg/kg, which is not suitable for long-term consumption by patients with peptic ulcer and gout.
  • the primary purpose of the present invention is to overcome the shortcomings and deficiencies in the prior art and provide a method for preparing low-purine soybean milk powder.
  • Another object of the present invention is to provide a low-purine soybean milk powder.
  • a method for preparing low-purine soy milk powder includes the following steps:
  • soybean milk fermentation medium dissolve the malt extract in soybean soybean milk, adjust the pH to 5.5-6.5, sterilize and cool to obtain a soybean milk fermentation medium;
  • Soymilk fermentation inoculate soymilk fermentation medium with purine-utilizing yeast for fermentation to obtain fermentation broth;
  • the malt extract in step (1) is preferably malt extract; more preferably food grade malt extract.
  • the soybean soy milk described in step (1) is preferably prepared by the following steps: washing with clear water, soaking soybeans, peeling, and draining water; adding water to the drained soybeans for beating and filtering to obtain soybean milk.
  • the soaking time is preferably 4-8h.
  • the rotation speed of the beating process is preferably 7000-8000 r/min.
  • the filtration is preferably passed through a 150-250 mesh sieve; more preferably, passed through a 200 mesh sieve.
  • the sterilization conditions described in step (1) are preferably sterilization at 110-130°C for 10-20 minutes; more preferably, sterilization at 121°C for 15 minutes.
  • the purine-utilizing yeast is preferably one or two of Blastobotrys adeninivorans and Candida utilis; more preferably, it is adenine-feeding Botrytis adeninivorans and Candida utilis.
  • the adenine-feeding Botrytis sphaeroides is preferably at least one of the adenine-feeding Botrytis spores CICC 33223 and CICC 33224 preserved in the China Industrial Microorganism Collection and Management Center.
  • the Candida utilis is preferably the Candida utilis CICC 32604 deposited at the China Industrial Microorganism Strain Collection and Management Center.
  • the purine-utilizing yeast is a yeast cell in the logarithmic growth phase or stable phase when it is inoculated into the soymilk fermentation medium; preferably, it is prepared by the following steps: Seed, inoculate in yeast culture medium, shake culture, obtain the culture solution containing yeast cells in logarithmic growth phase or stable phase; when the purine-utilizing yeast is adenine-feeding Botrytis yeast cells and prion pseudo In the case of silk yeast cell mixture, the adenine-feeding blastocystis slant strain and the Candida utilis cell slant strain are respectively inoculated in the same yeast medium and cultured with shaking to obtain the content in the logarithmic growth phase or stable phase A culture solution of a mixture of yeast cells.
  • the inoculation amount of the slant strain is preferably calculated by inoculating 1 inoculation ring lawn per 200 mL of yeast culture medium.
  • the temperature of the shaking culture is preferably 26-28°C.
  • the time of the shaking culture is preferably 18-22h.
  • the yeast culture medium is preferably prepared by the following steps: dissolving the malt extract in water, adjusting the pH to 5.5-6.5, and sterilizing to obtain the yeast culture medium.
  • the malt extract is preferably malt extract; more preferably, food grade malt extract.
  • the sterilization conditions are preferably 110-130°C for 10-20 minutes; more preferably 121°C for 15 minutes.
  • the inoculation amount of the purine-utilizable yeast is preferably 5% to 10% (v/v) equivalent to the volume of the soybean milk fermentation medium.
  • the fermentation described in step (2) is preferably aerobic fermentation.
  • the conditions of the aerobic fermentation are as follows: sterile air is introduced and stirred, the aeration ratio is 0.3-0.8 vvm, and the temperature is controlled at 26-28°C for 48-72 hours of fermentation.
  • the aeration ratio control is preferably as follows: during fermentation 0-24h, the aeration ratio is controlled to 0.3-0.5vvm; during 24-48h fermentation, the aeration ratio is controlled to be 0.5-0.8vvm; if the fermentation time is more than 48h, fermentation is 48h ⁇ During the end of fermentation, the aeration ratio is controlled at 0.5-0.8vvm.
  • the rotation speed of the stirring is preferably 200-240 r/min.
  • the filtration in step (3) is preferably ultrafiltration.
  • the molecular weight cut-off of the ultrafiltration membrane of the ultrafiltration is preferably 200-250kDa.
  • the concentration ratio of the ultrafiltration is preferably (8-12):1.
  • the concentration in step (3) is preferably concentrated by vacuum evaporation.
  • the temperature of the liquid concentrated by vacuum evaporation is preferably controlled at 46-50°C.
  • the degree of concentration in step (3) is preferably to control the concentration ratio (4-6):1.
  • the sterilization method described in step (3) is preferably ultra-high temperature instantaneous sterilization.
  • the ultra-high temperature instantaneous sterilization conditions are preferably: temperature 130-140°C, time 4-6s.
  • the homogenization in step (3) preferably adopts ultra-high pressure homogenization.
  • the pressure of the ultra-high pressure homogenization is preferably 30-40Mpa.
  • the drying described in step (3) is preferably spray drying.
  • a low-purine soybean milk powder is prepared by the above-mentioned preparation method of low-purine soybean milk powder.
  • the present invention has the following advantages and effects:
  • the invention uses mixed bacteria fermentation technology to prepare soybean milk powder.
  • the adenine-feeding Botrytis spores and Candida utilis can use some purines as fermentation carbon sources, and have certain metabolic decomposition capabilities for purines in soybean milk.
  • Mixed bacteria fermentation can greatly reduce the total purine content of soybean milk, and the removal efficiency is higher, generally reaching more than 90%.
  • the ultrafiltration membrane is used to trap the yeasts, and the nutrients of soy milk can pass through the ultrafiltration membrane, vacuum evaporation concentration, ultra-high temperature instantaneous sterilization, high pressure homogenization, and spray drying can obtain low-purine soybean milk powder.
  • the total purine content of low-purine soybean milk powder is reduced to below 140 mg/kg, which provides beneficial soybean milk powder for patients with peptic ulcer and gout.
  • Figure 1 is a flow chart of the production process of low-purine soybean milk powder 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. Unless otherwise specified, all reagents and materials used in the present invention are commercially available.
  • the adenine-feeding Botrytis spores CICC 33223, CICC 33224 and Candida utilis CICC 32604 were all purchased from the China Industrial Microbial Culture Collection and Management Center.
  • soybeans weigh 1.2 kg of soybeans, soak the soybeans in clean water for 6 hours, remove the hulls, and drain the water. Put the soybeans into the beater, add 16.8L of purified water, and beat at 7500r/min. Filter with a 200-mesh sieve and take the liquid under the sieve to obtain 18.1 L of soy milk. Put the soy milk into the fermentation tank, add 0.48kg of food-grade malt extract powder, stir to dissolve, adjust the pH to 6, sterilize at 121°C for 12 minutes, and cool for use.
  • yeast culture medium Divide the yeast culture medium into 7 1000mL Erlenmeyer flasks, each with a volume of 200mL, sterilize at 121°C for 15 minutes, after cooling, connect one loop (inoculation loop) of adenine-feeding Botrytis spore yeast CICC 33223 The lawn of the inclined plane strain, and the lawn with 1 loop (inoculation loop) of Candida utilis CICC 32604 inclined plane strain. Place the Erlenmeyer flask in a shaking incubator, control the temperature at 27°C, control the speed at 190r/min, and cultivate for 20 hours to obtain a mixed yeast culture solution.
  • the soy milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected as 200kDa, and the ultrafiltration concentration ratio was controlled to 10:1 to obtain 17.5L of permeate.
  • the total purine content of the prepared soybean milk powder was 131 mg/kg. Use 4 to 8 times the amount of water to make soy milk powder into soy milk, soy flavor is strong.
  • soybeans weigh 1.5 kg of soybeans, soak the soybeans in clean water for 4 hours, remove the hulls, and drain the water. Put the soybeans into the beater, add 18L of purified water, and beat at 8000r/min. Filter with a 200-mesh sieve and take the liquid under the sieve to obtain 20.6 L of soy milk. Put the soy milk into the fermentation tank, add 0.45kg of food-grade malt extract powder, stir to dissolve, adjust the pH to 5.5, sterilize at 121°C for 10 minutes, and cool for use.
  • the stirring speed is controlled at 240r/min, and the temperature is controlled at 26-28°C; the fermentation is 0-24h, and the aeration ratio is controlled at 0.3-0.5vvm; and the fermentation is 24-48h, and the aeration ratio is controlled at 0.5-0.8vvm.
  • the fermentation period is controlled to 48h.
  • the soybean milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected to be 250kDa, and the ultrafiltration concentration ratio was controlled to 12:1 to obtain 19.8L of permeate.
  • the total purine content of the prepared soybean milk powder is 118 mg/kg.
  • soybeans weigh 1.5 kg of soybeans, soak the soybeans in clean water for 8 hours, remove the hulls, and drain the water. Put the soybeans into the beater, add 24L of purified water, and beat at 7000r/min. Filter with a 200-mesh sieve and take the liquid under the sieve to obtain 26.4L of soy milk. Put the soy milk into the fermentation tank, add 0.75kg food grade malt extract powder, stir to dissolve, adjust the pH to 6.5, sterilize at 121°C for 15 minutes, and cool for use.
  • the stirring speed is controlled at 200r/min, the temperature is controlled at 26-28°C; the fermentation is 0-24h, and the aeration ratio is controlled at 0.3-0.5vvm; the fermentation is 24-72h, and the aeration ratio is controlled at 0.5-0.8vvm.
  • the fermentation period is controlled to 72h.
  • the soybean milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected to be 200kDa, and the ultrafiltration concentration ratio was controlled to 8:1 to obtain 25.4L of permeate.
  • the total purine content of the prepared soybean milk powder is 136 mg/kg.
  • soybeans weigh 1.2 kg of soybeans, soak the soybeans in clean water for 6 hours, remove the hulls, and drain the water. Put the soybeans into the beater, add 16.8L of purified water, and beat at 7500r/min. Filter with a 200-mesh sieve and take the liquid under the sieve to obtain 17.9L of soy milk. Put the soy milk into the fermentation tank, add 0.48kg of food-grade malt extract powder, stir to dissolve, adjust the pH to 6, sterilize at 121°C for 12 minutes, and cool for use.
  • yeast culture medium Divide the yeast culture medium into 7 1000mL Erlenmeyer flasks, each with a volume of 200mL, sterilize at 121°C for 15 minutes, after cooling, connect one loop (inoculation loop) of adenine-feeding Botrytis spore yeast CICC 33223 The lawn of the slant strain.
  • the triangular flask was placed in a shaking incubator, the temperature was controlled at 27°C, the rotation speed was controlled at 190r/min, and cultured for 20 hours to obtain adenine-feeding budding Botrytis yeast culture solution.
  • the soybean milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected to be 200kDa, and the ultrafiltration concentration ratio was controlled to 10:1 to obtain 17.3L of permeate.
  • the total purine content of the prepared soybean milk powder is 1014 mg/kg.
  • soybeans weigh 1.2 kg of soybeans, soak the soybeans in clean water for 6 hours, remove the hulls, and drain the water. Put the soybeans into the beater, add 16.8L of purified water, and beat at 7500r/min. Filter with a 200-mesh sieve and take the liquid under the sieve to obtain 18L of soy milk. Put the soy milk into the fermentation tank, add 0.48kg of food-grade malt extract powder, stir to dissolve, adjust the pH to 6, sterilize at 121°C for 12 minutes, and cool for use.
  • yeast culture medium Divide the yeast culture medium into 7 1000mL Erlenmeyer flasks, each with a volume of 200mL, sterilize at 121°C for 15 minutes, after cooling, connect 1 loop (inoculation loop) adenine-feeding botrytis yeast CICC 33224 The lawn of the slant strain.
  • the triangular flask was placed in a shaking incubator, the temperature was controlled at 27°C, the rotation speed was controlled at 190r/min, and cultured for 20 hours to obtain adenine-feeding budding Botrytis yeast culture solution.
  • the soybean milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected to be 200kDa, and the ultrafiltration concentration ratio was controlled to 10:1 to obtain 17.4L of permeate.
  • the total purine content of the prepared soybean milk powder is 967mg/kg.
  • soybeans weigh 1.2 kg of soybeans, soak the soybeans in clean water for 6 hours, remove the hulls, and drain the water. Put the soybeans into the beater, add 16.8L of purified water, and beat at 7500r/min. Filter with a 200-mesh sieve and take the liquid under the sieve to obtain 18.1 L of soy milk. Put the soy milk into the fermentation tank, add 0.48kg of food-grade malt extract powder, stir to dissolve, adjust the pH to 6, sterilize at 121°C for 12 minutes, and cool for use.
  • the stirring speed is controlled at 220r/min, and the temperature is controlled at 26-28°C; the fermentation is 0-24h, and the aeration ratio is controlled at 0.3-0.5vvm; the fermentation is 24-60h, and the aeration ratio is controlled at 0.5-0.8vvm.
  • the fermentation cycle is controlled to 60h.
  • the soy milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected as 200kDa, and the ultrafiltration concentration ratio was controlled to 10:1 to obtain 17.5L of permeate.
  • the total purine content of the prepared soybean milk powder is 2165 mg/kg.
  • the soybean milk was filtered with an ultrafiltration machine, the molecular weight cut-off of the ultrafiltration membrane was selected to be 200kDa, and the ultrafiltration concentration ratio was controlled to 10:1 to obtain 16.38L of permeate.
  • the total purine content of the prepared soybean milk powder is 2574mg/kg.

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Abstract

一种低嘌呤豆浆粉及其制作方法,首先,将豆浆与麦芽提取物配制成发酵培养基,通过能利用嘌呤的酵母进行发酵,分解豆浆中的嘌呤类物质;然后,利用过滤、浓缩、灭菌、均质、干燥等技术,对发酵液进行处理,获得低嘌呤豆浆粉。

Description

一种低嘌呤豆浆粉及其制作方法 技术领域
本发明属于食品加工技术领域,特别涉及一种低嘌呤豆浆粉及其制作方法。
背景技术
豆浆是大豆经浸泡、磨浆、过滤、煮沸等步骤制成的饮品,其中的蛋白质、脂肪、矿物质和维生素等营养物质含量丰富,还含有大豆异黄酮、卵磷脂等保健功能物质,素有“植物奶”美誉。将豆浆制成豆浆粉,可以满足消费者在不同场合的冲饮需求。但是,无论是现磨的豆浆,还是利用豆浆粉冲调的豆浆,总嘌呤含量都很高。目前,绝大多数市售豆浆粉(不含糖类的商品)的总嘌呤含量为1800~2600mg/kg,不适宜消化性溃疡和痛风病患者长期食用。降低豆浆的嘌呤类物质,减小豆浆食用的禁忌,是豆浆制作的重要方向之一。国内外研究人员已在降低豆浆嘌呤类物质的技术领域进行了一些研究,主要是利用一些活性炭、壳聚糖等吸附剂去除豆浆中的嘌呤类物质,但是去除率仍较低,一般为50%左右。
发明内容
本发明的首要目的在于克服现有技术中的缺点与不足,提供一种低嘌呤豆浆粉的制作方法。
本发明的另一目的在于提供一种低嘌呤豆浆粉。
本发明的目的通过下述技术方案实现:
一种低嘌呤豆浆粉的制作方法,包括如下步骤:
(1)豆浆发酵培养基的配制:将麦芽提取物溶于大豆豆浆中,调节pH至5.5~6.5,灭菌,冷却,得到豆浆发酵培养基;
(2)豆浆的发酵:在豆浆发酵培养基中接种能利用嘌呤的酵母进行发酵,得到发酵液;
(3)发酵液的干燥处理:将所得发酵液经过滤,浓缩,灭菌,均质,干燥,获得低嘌呤豆浆粉。
步骤(1)中所述的麦芽提取物优选为麦芽浸粉;更优选为食品级麦芽浸粉。
步骤(1)中所述的麦芽提取物的用量优选按麦芽提取物:大豆=(0.3~0.5):1的质量比计算。
步骤(1)中所述的大豆豆浆优选通过如下步骤制备得到:用清水清洗、浸泡大豆,脱皮,沥干水分;在沥干后的大豆中再加入水打浆,过滤,得到豆浆。
所述的浸泡的时间优选为4~8h。
所述的打浆过程的加水量优选按水:大豆=(12~16):1的质量比计算。
所述的打浆过程的转速优选为7000~8000r/min。
所述的过滤优选为过150~250目筛;更优选为过200目筛。
步骤(1)中所述的灭菌的条件优选为于110~130℃下灭菌10~20min;更优选为于121℃下灭菌15min。
所述的能利用嘌呤的酵母优选为食腺嘌呤芽生葡萄孢酵母(Blastobotrys adeninivorans)和产朊假丝酵母(Candida utilis)中的一种或两种;更优选为食腺嘌呤芽生葡萄孢酵母和产朊假丝酵母按1:1配比得到的酵母细胞混合物。
所述的食腺嘌呤芽生葡萄孢酵母优选为保藏于中国工业微生物菌种保藏管理中心的食腺嘌呤芽生葡萄孢酵母CICC 33223和CICC 33224中的至少一种。
所述的产朊假丝酵母优选为保藏于中国工业微生物菌种保藏管理中心的产朊假丝酵母CICC 32604。
所述的能利用嘌呤的酵母在接种到所述的豆浆发酵培养基时是处于对数生长期或稳定期的酵母细胞;优选通过如下步骤制备得到:将所述的能利用嘌呤的酵母斜面菌种,接种于酵母培养基,振荡培养,得到含处于对数生长期或稳定期的酵母细胞的培养液;当所述的能利用嘌呤的酵母为食腺嘌呤芽生葡萄孢酵母细胞和产朊假丝酵母细胞混合物时,是将食腺嘌呤芽生葡萄孢酵母斜面菌种和产朊假丝酵母细胞斜面菌种分别接种于同一酵母培养基,振荡培养,得到含处于对数生长期或稳定期的酵母细胞混合物的培养液。
所述的斜面菌种的接种量优选按每200mL酵母培养基接种1接种环菌苔计算。
所述的振荡培养的温度优选为26~28℃。
所述的振荡培养的时间优选为18~22h。
所述的酵母培养基优选为通过如下步骤配制得到:将麦芽提取物溶于水,调节pH至5.5~6.5,灭菌,得到酵母培养基。
所述的麦芽提取物优选为麦芽浸粉;更优选为食品级麦芽浸粉。
所述的麦芽提取物的加入量优选为按麦芽提取物:水=1:(20~24)的质量比计算。
所述的灭菌的条件优选为于110~130℃下灭菌10~20min;更优选为于121℃下灭菌15min。
所述的能利用嘌呤的酵母的接种量优选按相当于豆浆发酵培养基体积的5%~10%(v/v)计。
步骤(2)中所述的发酵优选为好氧发酵。
所述的好氧发酵的条件为:通入无菌空气并加以搅拌,通气比为0.3~0.8vvm,控制温度为26~28℃发酵48~72h。
所述的通气比的控制优选如下:发酵0~24h期间,通气比控制为0.3~0.5vvm;发酵 24~48h期间,通气比控制为0.5~0.8vvm;如果发酵时间在48h以上,发酵48h~发酵结束期间,通气比控制为0.5~0.8vvm。
所述的搅拌的转速优选为200~240r/min。
步骤(3)中所述的过滤优选为超滤。
所述的超滤的超滤膜截留分子量优选为200~250kDa。
所述的超滤的浓缩比优选为(8~12):1。
步骤(3)中所述的浓缩优选为采用真空蒸发浓缩。
所述的真空蒸发浓缩的液体温度优选控制在46~50℃。
步骤(3)中所述的浓缩的程度优选为控制浓度比(4~6):1。
步骤(3)中所述的灭菌方法优选为超高温瞬时灭菌。
所述的超高温瞬时灭菌条件优选为:温度130~140℃,时间4~6s。
步骤(3)中所述的均质优选采用超高压均质。
所述的超高压均质的压力优选为30~40Mpa。
步骤(3)中所述的干燥优选为喷雾干燥。
一种低嘌呤豆浆粉,通过上述的低嘌呤豆浆粉的制作方法制成。
本发明相对于现有技术具有如下的优点及效果:
本发明利用混合菌发酵技术制备豆浆粉,所用食腺嘌呤芽生葡萄孢酵母和产朊假丝酵母能够以一些嘌呤类物质为发酵碳源,对豆浆中的嘌呤类物质分别具有一定的代谢分解能力,混合菌发酵能够使豆浆的总嘌呤含量大幅度降低,去除效率较高,一般达到90%以上。发酵后,利用超滤膜截留酵母菌,豆浆的营养成分可以透过超滤膜,经真空蒸发浓缩、超高温瞬时灭菌、高压均质、喷雾干燥,可获得低嘌呤的豆浆粉。低嘌呤豆浆粉的总嘌呤含量降低至140mg/kg以下,为消化性溃疡和痛风病患者提供了有益的豆浆粉。
附图说明
图1为本发明低嘌呤豆浆粉的制作工艺流程图。
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。
本发明实施例中的食腺嘌呤芽生葡萄孢酵母CICC 33223、CICC 33224和产朊假丝酵母CICC 32604均购于中国工业微生物菌种保藏管理中心。
实施例1
一种低嘌呤豆浆粉的制作过程,如图1所示,具体如下:
(1)豆浆发酵培养基的配制
称取1.2kg大豆,用清水浸泡大豆6h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加入16.8L纯净水,于7500r/min的条件下打浆。用200目筛过滤,取筛下液体,获得18.1L豆浆。将豆浆放入发酵罐内,加入0.48kg食品级麦芽浸粉,搅拌溶解,调节pH至6,于121℃下灭菌12min,冷却后备用。
(2)豆浆的发酵
称取63.6g食品级麦芽浸粉,加入1400mL水,搅拌溶解,调节pH至6,配制成为酵母培养基。将酵母培养基分装至7个1000mL三角瓶,每瓶装液量为200mL,于121℃下灭菌15min,冷却后,每瓶接入1环(接种环)食腺嘌呤芽生葡萄孢酵母CICC 33223斜面菌种的菌苔,以及接入1环(接种环)产朊假丝酵母CICC 32604斜面菌种的菌苔。将三角瓶置于振荡培养箱中,温度控制为27℃,转速控制为190r/min,培养20h,获得混合酵母菌培养液。
将1360mL混合酵母菌培养液接入发酵罐,启动搅拌,并通入无菌空气。在发酵过程中,搅拌转速控制为220r/min,温度控制为26~28℃;发酵0~24h,通气比控制为0.3~0.5vvm;发酵24~60h,通气比控制为0.5~0.8vvm。发酵周期控制为60h。
(3)发酵液的干燥处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为200kDa,超滤浓缩比控制为10:1,获得17.5L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至3.5L,然后将浓缩液于135℃的条件下灭菌5s,再在35Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.72kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为131mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
实施例2
(1)豆浆发酵培养基的配制
称取1.5kg大豆,用清水浸泡大豆4h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加入18L纯净水,于8000r/min的条件下打浆。用200目筛过滤,取筛下液体,获得20.6L豆浆。将豆浆放入发酵罐内,加入0.45kg食品级麦芽浸粉,搅拌溶解,调节pH至5.5,于121℃下灭菌10min,冷却后备用。
(2)豆浆的发酵
称取60g食品级麦芽浸粉,加入1200mL水,搅拌溶解,调节pH至5.5,配制成为酵母培养基。将酵母培养基分装至6个1000mL三角瓶,每瓶装液量为200mL,于121℃下灭菌15min,冷却后,每瓶接入1环(接种环)食腺嘌呤芽生葡萄孢酵母CICC 33224斜面菌种的菌苔,以及接入1环(接种环)产朊假丝酵母CICC 32604斜面菌种的菌苔。将三角瓶置于振荡培养箱中,温度控制为26℃,转速控制为180r/min,培养22h,获得混合酵母菌培养液。
将1030mL混合酵母菌培养液接入发酵罐,启动搅拌,并通入无菌空气。在发酵过程中,搅拌转速控制为240r/min,温度控制为26~28℃;发酵0~24h,通气比控制为0.3~0.5vvm;发酵24~48h,通气比控制为0.5~0.8vvm。发酵周期控制为48h。
(3)发酵液的干燥处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为250kDa,超滤浓缩比控制为12:1,获得19.8L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至4.95L,然后将浓缩液于130℃的条件下灭菌6s,再在30Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.91kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为118mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
实施例3
(1)豆浆发酵培养基的配制
称取1.5kg大豆,用清水浸泡大豆8h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加入24L纯净水,于7000r/min的条件下打浆。用200目筛过滤,取筛下液体,获得26.4L豆浆。将豆浆放入发酵罐内,加入0.75kg食品级麦芽浸粉,搅拌溶解,调节pH至6.5,于121℃下灭菌15min,冷却后备用。
(2)豆浆的发酵
称取116.7g食品级麦芽浸粉,加入2800mL水,搅拌溶解,调节pH至6.5,配制成为酵母培养基。将酵母培养基分装至14个1000mL三角瓶,每瓶装液量为200mL,于121℃下灭菌15min,冷却后,每瓶接入1环(接种环)食腺嘌呤芽生葡萄孢酵母CICC 33223斜面菌种的菌苔,以及接入1环(接种环)产朊假丝酵母CICC 32604斜面菌种的菌苔。将三角瓶置于振荡培养箱中,温度控制为28℃,转速控制为200r/min,培养18h,获得混合酵母菌培养液。
将2640mL混合酵母菌培养液接入发酵罐,启动搅拌,并通入无菌空气。在发酵过程中,搅拌转速控制为200r/min,温度控制为26~28℃;发酵0~24h,通气比控制为0.3~0.5vvm;发酵24~72h,通气比控制为0.5~0.8vvm。发酵周期控制为72h。
(3)发酵液的干燥处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为200kDa,超滤浓缩比控制为8:1,获得25.4L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至4.23L,然后将浓缩液于140℃的条件下灭菌4s,再在40Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.93kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为136mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
实施例4
(1)豆浆发酵培养基的配制
称取1.2kg大豆,用清水浸泡大豆6h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加入16.8L纯净水,于7500r/min的条件下打浆。用200目筛过滤,取筛下液体,获得17.9L豆浆。将豆浆放入发酵罐内,加入0.48kg食品级麦芽浸粉,搅拌溶解,调节pH至6,于121℃下灭菌12min,冷却后备用。
(2)豆浆的发酵
称取63.6g食品级麦芽浸粉,加入1400mL水,搅拌溶解,调节pH至6,配制成为酵母培养基。将酵母培养基分装至7个1000mL三角瓶,每瓶装液量为200mL,于121℃下灭菌15min,冷却后,每瓶接入1环(接种环)食腺嘌呤芽生葡萄孢酵母CICC 33223斜面菌种的菌苔。将三角瓶置于振荡培养箱中,温度控制为27℃,转速控制为190r/min,培养20h,获得食腺嘌呤芽生葡萄孢酵母培养液。
将1360mL食腺嘌呤芽生葡萄孢酵母培养液接入发酵罐,启动搅拌,并通入无菌空气。在发酵过程中,搅拌转速控制为220r/min,温度控制为26~28℃;发酵0~24h,通气比控制为0.3~0.5vvm;发酵24~60h,通气比控制为0.5~0.8vvm。发酵周期控制为60h。
(3)发酵液的干燥处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为200kDa,超滤浓缩比控制为10:1,获得17.3L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至3.46L,然后将浓缩液于135℃的条件下灭菌5s,再在35Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.73kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为1014mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
实施例5
(1)豆浆发酵培养基的配制
称取1.2kg大豆,用清水浸泡大豆6h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加入16.8L纯净水,于7500r/min的条件下打浆。用200目筛过滤,取筛下液体,获得18L豆浆。将豆浆放入发酵罐内,加入0.48kg食品级麦芽浸粉,搅拌溶解,调节pH至6,于121℃下灭菌12min,冷却后备用。
(2)豆浆的发酵
称取63.6g食品级麦芽浸粉,加入1400mL水,搅拌溶解,调节pH至6,配制成为酵母培养基。将酵母培养基分装至7个1000mL三角瓶,每瓶装液量为200mL,于121℃下灭菌15min,冷却后,每瓶接入1环(接种环)食腺嘌呤芽生葡萄孢酵母CICC 33224斜面菌种的菌苔。将三角瓶置于振荡培养箱中,温度控制为27℃,转速控制为190r/min,培养20h,获得食腺嘌呤芽生葡萄孢酵母培养液。
将1360mL食腺嘌呤芽生葡萄孢酵母培养液接入发酵罐,启动搅拌,并通入无菌空气。 在发酵过程中,搅拌转速控制为220r/min,温度控制为26~28℃;发酵0~24h,通气比控制为0.3~0.5vvm;发酵24~60h,通气比控制为0.5~0.8vvm。发酵周期控制为60h。
(3)发酵液的干燥处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为200kDa,超滤浓缩比控制为10:1,获得17.4L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至3.48L,然后将浓缩液于135℃的条件下灭菌5s,再在35Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.72kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为967mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
实施例6
(1)豆浆发酵培养基的配制
称取1.2kg大豆,用清水浸泡大豆6h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加入16.8L纯净水,于7500r/min的条件下打浆。用200目筛过滤,取筛下液体,获得18.1L豆浆。将豆浆放入发酵罐内,加入0.48kg食品级麦芽浸粉,搅拌溶解,调节pH至6,于121℃下灭菌12min,冷却后备用。
(2)豆浆的发酵
称取63.6g食品级麦芽浸粉,加入1400mL水,搅拌溶解,调节pH至6,配制成为酵母培养基。将酵母培养基分装至7个1000mL三角瓶,每瓶装液量为200mL,于121℃下灭菌15min,冷却后,每瓶接入1环(接种环)产朊假丝酵母CICC 32604斜面菌种的菌苔。将三角瓶置于振荡培养箱中,温度控制为27℃,转速控制为190r/min,培养20h,获得产朊假丝酵母培养液。
将1360mL产朊假丝酵母培养液接入发酵罐,启动搅拌,并通入无菌空气。在发酵过程中,搅拌转速控制为220r/min,温度控制为26~28℃;发酵0~24h,通气比控制为0.3~0.5vvm;发酵24~60h,通气比控制为0.5~0.8vvm。发酵周期控制为60h。
(3)发酵液的干燥处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为200kDa,超滤浓缩比控制为10:1,获得17.5L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至3.5L,然后将浓缩液于135℃的条件下灭菌5s,再在35Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.73kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为2165mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
对比例1
(1)豆浆制取
称取1.2kg大豆,用清水浸泡大豆6h,脱除豆皮,沥干水分。将黄豆放入打浆机中,加 入16.8L纯净水,于7500r/min的条件下打浆。用200目筛过滤,取筛下液体,获得18.2L豆浆。
(2)豆浆的后处理
用超滤机对豆浆进行过滤,超滤膜的截留分子量选择为200kDa,超滤浓缩比控制为10:1,获得16.38L透过液。利用真空蒸发器在在46~50℃下将透过液浓缩至3.28L,然后将浓缩液于135℃的条件下灭菌5s,再在35Mpa的压力下进行超高压均质,最后经喷雾干燥,获得0.804kg豆浆粉。经检测,所制得豆浆粉的总嘌呤含量为2574mg/kg。用4~8倍的水将豆浆粉冲调成豆浆,豆香风味浓郁。
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。

Claims (10)

  1. 一种低嘌呤豆浆粉的制作方法,其特征在于:包括如下步骤:
    (1)豆浆发酵培养基的配制:将麦芽提取物溶于大豆豆浆中,调节pH至5.5~6.5,灭菌,冷却,得到豆浆发酵培养基;
    (2)豆浆的发酵:在豆浆发酵培养基中接种能利用嘌呤的酵母进行发酵,得到发酵液;
    (3)发酵液的干燥处理:将所得发酵液经过滤,浓缩,灭菌,均质,干燥,获得低嘌呤豆浆粉;
    所述的能利用嘌呤的酵母为食腺嘌呤芽生葡萄孢酵母(Blastobotrys adeninivorans)和产朊假丝酵母(Candida utilis)中的一种或两种。
  2. 根据权利要求1所述的低嘌呤豆浆粉的制作方法,其特征在于:
    所述的食腺嘌呤芽生葡萄孢酵母为食腺嘌呤芽生葡萄孢酵母CICC 33223和食腺嘌呤芽生葡萄孢酵母CICC 33224中的至少一种;
    所述的产朊假丝酵母为产朊假丝酵母CICC 32604。
  3. 根据权利要求1或2所述的低嘌呤豆浆粉的制作方法,其特征在于:
    步骤(2)中所述的能利用嘌呤的酵母为处于对数生长期或稳定期的酵母细胞;
    所述的处于对数生长期或稳定期的酵母细胞通过如下步骤制备得到:将能利用嘌呤的酵母斜面菌种,接种于同一酵母培养基,振荡培养,得到含处于对数生长期或稳定期的酵母细胞的培养液;当所述的能利用嘌呤的酵母为食腺嘌呤芽生葡萄孢酵母细胞和产朊假丝酵母细胞混合物时,是将食腺嘌呤芽生葡萄孢酵母斜面菌种和产朊假丝酵母细胞斜面菌种分别接种于同一酵母培养基,振荡培养,得到含处于对数生长期或稳定期的酵母细胞混合物的培养液;
    步骤(2)中所述的发酵为好氧发酵。
  4. 根据权利要求3所述的低嘌呤豆浆粉的制作方法,其特征在于:
    所述的斜面菌种的接种量均按每200mL酵母培养基接种1接种环菌苔计;
    所述的振荡培养的温度为26~28℃;
    所述的振荡培养的时间为18~22h;
    所述的酵母培养基通过如下步骤配置得到:将麦芽提取物溶于水,调节pH至5.5~6.5,灭菌,得到酵母培养基;
    所述的好氧发酵的条件为:通入无菌空气并加以搅拌,通气比为0.3~0.8vvm,控制温度为26~28℃发酵48~72h。
  5. 根据权利要求4所述的低嘌呤豆浆粉的制作方法,其特征在于:
    所述的麦芽提取物为麦芽浸粉;
    所述的麦芽提取物的加入量按麦芽提取物:水=1:(20~24)的质量比计算;
    所述的酵母培养基配置过程中的灭菌的条件为于110~130℃下灭菌10~20min;
    所述的搅拌的转速为200~240r/min;
    所述的通气比的控制如下:发酵0~24h期间,通气比控制为0.3~0.5vvm;发酵24~48h期间,通气比控制为0.5~0.8vvm;如果发酵时间在48h以上,发酵48h~发酵结束期间,通气比控制为0.5~0.8vvm。
  6. 根据权利要求1或2所述的低嘌呤豆浆粉的制作方法,其特征在于:
    步骤(1)中所述的麦芽提取物为麦芽浸粉;
    步骤(1)中所述的麦芽提取物的用量按麦芽提取物:大豆=(0.3~0.5):1的质量比计算;
    步骤(1)中所述的大豆豆浆通过如下步骤制备得到:用清水清洗、浸泡大豆,脱皮,沥干水分;在沥干后的大豆中再加入水打浆,过滤,得到豆浆;
    步骤(1)中所述的灭菌的条件为于110~130℃下灭菌10~20min。
  7. 根据权利要求6所述的低嘌呤豆浆粉的制作方法,其特征在于:
    所述的麦芽提取物为食品级麦芽浸粉;
    所述的浸泡的时间为4~8h;
    所述的打浆过程的加水量按水:大豆=(12~16):1的质量比计算;
    所述的打浆过程的转速为7000~8000r/min;
    所述的过滤为过150~250目筛;
    步骤(1)中所述的灭菌的条件为于121℃下灭菌15min。
  8. 根据权利要求1或2所述的低嘌呤豆浆粉的制作方法,其特征在于:
    步骤(3)中所述的过滤为超滤;
    步骤(3)中所述的浓缩为采用真空蒸发浓缩;
    步骤(3)中所述的浓缩的程度为控制浓度比(4~6):1;
    步骤(3)中所述的灭菌方法为超高温瞬时灭菌;
    步骤(3)中所述的均质采用超高压均质;
    步骤(3)中所述的干燥为喷雾干燥。
  9. 根据权利要求8所述的低嘌呤豆浆粉的制作方法,其特征在于:
    所述的超滤的超滤膜截留分子量为200~250kDa;
    所述的超滤的浓缩比为(8~12):1;
    所述的真空蒸发浓缩的液体温度控制在46~50℃;
    所述的超高温瞬时灭菌条件为:温度130~140℃,时间4~6s;
    所述的超高压均质的压力为30~40Mpa。
  10. 一种低嘌呤豆浆粉,其特征在于:通过权利要求1~9任一项所述的低嘌呤豆浆粉的制作方法制成。
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