CN111607621A - Yeast capable of producing rose fragrance and application of yeast in Lingwu jujube enzyme - Google Patents

Yeast capable of producing rose fragrance and application of yeast in Lingwu jujube enzyme Download PDF

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CN111607621A
CN111607621A CN202010396033.8A CN202010396033A CN111607621A CN 111607621 A CN111607621 A CN 111607621A CN 202010396033 A CN202010396033 A CN 202010396033A CN 111607621 A CN111607621 A CN 111607621A
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yeast
jujube
lingwu
producing
aroma
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许育衔
郁小宁
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Jingye Yinchuan Biotechnology Co ltd
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Jingye Yinchuan Biotechnology Co ltd
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    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
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    • C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/78—Hansenula

Abstract

The invention belongs to the technical field of food processing, and particularly relates to a yeast for producing rose fragrance and application thereof in Lingwu jujube enzyme, wherein the yeast comprises the following components in parts by weight: the aroma-producing yeast is separated from the Lingwu jujube juice and is a strain of yeast with local characteristics. The aroma-producing yeast is Hansenula polymorpha JYY015 of grape juice, and the preservation number is CGMCC NO. 18969; the aroma components are detected by culturing for 48h in YPD medium, the highest relative content is phenethylacetate with rose fragrance, and the relative content reaches 29.8732%. The aroma-producing yeast is added at the initial stage of Lingwu long jujube fermentation, the inoculation amount is 3% -5%, and the fermentation is carried out for 48-72h at 28-30 ℃, so that the fermentation liquor not only has jujube aroma, but also has unique fermentation aroma. The strain grows rapidly, the culture condition is simple, the ester production efficiency is high, and the strain is easy to be used for industrial production.

Description

Yeast capable of producing rose fragrance and application of yeast in Lingwu jujube enzyme
Technical Field
The invention relates to the technical field of food processing, in particular to a yeast capable of producing rose fragrance and application of the yeast in Lingwu date enzyme.
Background
Lingwu long jujube, Rhamnaceae, alias Maya jujube, is a good characteristic economic variety in the autonomous region of Ningxia Hui nationality, and has a long cultivation history. The natural conditions of Ningxia Lingwu city make Lingwu jujube produce large fruit, round long handle, delicious and juicy fruit, crisp taste, thin peel, white and green pulp, rich nutrients and high vitamin content. Since 2003, the Lingwu long jujube has been developed rapidly and successfully promotes the economic development of Lingwu city, and has good economic benefit, social benefit and ecological benefit.
The Lingwu long jujube is similar to other fresh eating jujubes of various excellent varieties, and due to the physiological characteristics of compact pulp texture, low water content, vigorous anaerobic respiration and metabolism and the like, the jujube is very easy to be alcoholized and softened after being picked, and various substances in the jujube are subjected to different chemical changes. The Lingwu long jujube has high nutritive value, contains less common nutrients such as saccharides, amino acids, vitamins and minerals and special functional components with high physiological activity, such as cAMP (cyclic adenosine monophosphate), red jujube polysaccharide, saponin, flavone, alkaloid and the like, and the functional components enable the Lingwu long jujube to have various health-care functions such as bacteriostasis, liver protection, lipid reduction, organism immunity enhancement and the like.
The fermentation is a modern processing method of the jujube fruit, compared with the traditional fresh jujube juice, the method has the advantages that the complex components in the jujube fruit can be decomposed into simple substances by using microorganisms through the fermentation, the absorption of nutrient substances of the jujube fruit by a human body is facilitated, and the method has double health care values of the jujube and microbial metabolites. From the fragrance perspective, the fermented jujube juice not only contains the basic jujube fragrance, but also has pleasant sweet brewing fragrance.
Disclosure of Invention
This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, certain simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.
The invention aims to provide a yeast for producing rose fragrance with local characteristics, which is used in Lingwu jujube ferment, and also provides a method for applying the yeast for producing rose fragrance in Lingwu jujube ferment. Solves the problem of insufficient fermentation aroma of the long jujube enzyme, and improves the flavor characteristics and the nutritional value of the long jujube enzyme. And the yeast is used as the main bacterium for early fermentation by utilizing the characteristics of rapid growth, early time for entering the death phase and high efficiency of metabolic fragrance production of the yeast in the early fermentation stage.
To solve the above technical problem, according to an aspect of the present invention, the present invention provides the following technical solutions:
the invention discloses an aroma-producing bacterium JYY015, which has the colony morphology characteristics that: the strain is streaked and inoculated on YPD (glucose 2%, yeast powder 1%, peptone 2% and agar 2%) culture medium, and cultured at 28 deg.C for 2d, the diameter of colony is 3-5mm, the colony is circular and raised, its color is milk white, opaque, colony edge is smooth and non-irregular, and the microscopical thallus is lemon-type and bowling-ball-type.
Further, the aroma-producing bacteria provided by the invention are Hansenula polymorpha. The strain is preserved in China general microbiological culture Collection center (CGMCC) at 19.11.2019 at the location of No. 3 Hospital No.1 Chen West Lu of the sunward area in Beijing. The preservation number is CGMCC No.18969, the Latin name is Hanseniasps ra uvarum, and the name is Hansenula polymorpha JYY 015.
And (3) sequencing 18S region rDNA of the yeast strain, wherein the sequencing result is the nucleotide sequence shown in SEQ.ID.NO1.
Further, the yeast is cultured in YPD liquid for 48h, volatile components in the culture liquid are obtained by a rotary evaporator, and the aroma components are analyzed by gas chromatography-mass spectrometry, so that the bacterium is determined to have the highest content of the volatile component of phenethyl acetate in the YPD culture medium, and the content of the volatile component is 29.87%. The ingredient odor is described as a rose scent.
An application of the aroma-producing yeast in Lingwu long jujube enzyme:
the application of the aroma-producing yeast in Lingwu jujube ferment comprises the following steps:
s1: pretreatment of raw materials: purchasing Lingwu long jujube, cleaning, sterilizing, adding a certain proportion of sterile water, crushing and homogenizing. Then filtering with 30 mesh filter sieve to remove residue, avoiding the influence of fruit residue on taste, and pasteurizing the filtrate.
S2: adding auxiliary materials: adding 2% of sterilized sucrose, 10% of sterilized maltitol and 3% of sterilized inulin into the filtrate of the jujube juice.
S3: and (3) bacteria planting: inoculating Hansenula polymorpha JYY015 to grape juice, fermenting at 28-30 deg.C for 48-72 h.
S4: secondary bacterium planting: inoculating acetic acid bacteria and lactobacillus, fermenting at 30-33 deg.C for 24 hr, sealing, and fermenting under anaerobic condition for 4-7 days with pH of 3.2-3.6.
Further, a preferred embodiment of the present invention: in step S1, the raw material is preferably fresh, crisp, tender, rotten and pest-free Lingwu long jujube. Cleaning with ozone water twice, fumigating with ozone for 30min to eliminate surface bacteria as much as possible and avoid pathogenic bacteria residue. When the homogenate was broken, 2 times by weight of sterile water was added.
Further, a preferred embodiment of the present invention: in step S1, the filtered filtrate is sterilized, preferably by pasteurization (75-85 deg.C, incubation for 15-16S). The sterilization time is short, and the working efficiency is high. Not only can kill pathogenic bacteria, but also can not cause more nutrient loss due to too high temperature.
Further, a preferred embodiment of the present invention: in step S2, the auxiliary materials are added, preferably sucrose with purity of more than 99%, maltitol and inulin as auxiliary materials. The maltitol is not easy to be utilized by human body, has low calorie and is a safe sugar substitute product. Inulin is a natural water-soluble dietary fiber, hardly hydrolyzed and digested by gastric acid, and is used as a prebiotic and utilized only in the colon by beneficial microorganisms, thereby improving the intestinal environment. The auxiliary materials are fumigated by ozone for 30min for disinfection, so that no pathogenic bacteria are brought in during the addition process.
Further, a preferred embodiment of the present invention: in step S3, the strain is planted, preferably, a single strain Hansenula polymorpha JYY015 is selected, and the inoculation amount is 3% -5%. The ganoderma lucidum and Wu date pulp is fermented, so that the jujube pulp has stronger fragrance and has the unique fragrance of yeast. The yeast has rapid growth and vigorous metabolism, decomposes complex macromolecules in the fermentation substrate into micromolecules, and increases the nutritive value.
Further, a preferred embodiment of the present invention: in the step S4, secondary bacteria planting is carried out, and the secondary bacteria planting is inoculated into the fermentation substrate according to the proportion of 1: 1 of acetic acid bacteria and lactic acid bacteria and the inoculation amount of 5-6%. After secondary bacteria planting, introducing sterile air filtered by the filter membrane for 24 hours, wherein the dissolved oxygen is 4-12ppm/L, and the purpose is to provide an aerobic environment for aerobic acetic acid bacteria, so that the acetic acid bacteria quickly utilize the ethanol of the metabolite of the early-stage yeast to convert the ethanol into acetic acid, and ensure that the fermentation liquor has no ethanol residue. And (3) after 24 hours of ventilation, stopping ventilation, and entering an anaerobic fermentation stage, wherein the fermentation temperature is 30-33 ℃, and the lactobacillus becomes an absolute dominant strain after fermentation for 4-7 days. The invention utilizes the mutual beneficial symbiosis of the saccharomycetes, the lactic acid bacteria and the acetic acid bacteria, improves the utilization rate of raw materials and promotes the comprehensive enzyme activity of the product.
Compared with the prior art:
1. through the mixed fermentation of saccharomycetes, lactic acid bacteria and acetic acid bacteria, cAMP, rutin and other substances with higher health care value contained in the Lingwu long jujube are converted and hydrolyzed in the fermentation process, so that the solubility and the absorptivity of the Lingwu long jujube are greatly improved, and the nutritional value of the Lingwu long jujube is relatively improved.
2. The yeast selected for fermenting Lingwu long jujube is Hansenula polymorpha JYY015 of grape juice, so that the fresh jujube has strong fruit fragrance, unique fermentation fragrance is given, pleasant flower fragrance is accompanied, the taste is mellow and harmonious, besides, JYY015 is carried out along with the fermentation time, the death is rapid, no viable thallus exists in the later fermentation period, the problems of ethanol residue in the fermentation liquid, product flatulence and the like are solved, and the product quality is stable.
Through fermentation, the total sugar of the Lingwu long jujube enzyme is reduced, the auxiliary materials are mainly maltitol, and the functional sugar alcohol is a safe sugar substitute product, so that the purpose of reducing blood sugar is achieved, the comfortable taste is ensured, and the nutritional value is improved.
Drawings
FIG. 1 is a schematic representation of a JYY015 microscope (10X 100);
FIG. 2 is a colony morphology of JYY 015;
fig. 3 is a graph of JYY015 growth curves.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without any inventive step, shall fall within the scope of the present invention.
In the examples, it is to be noted that:
(1) preparing yeast, lactic acid bacteria and acetic acid bacteria seed liquid:
lactic acid bacteria seed liquid: selecting milk powder culture medium (10% milk powder, 2% glucose), sterilizing in 90 deg.C water bath for 20min, inoculating lactobacillus bacterial liquid into milk powder culture medium at 5% inoculum size, and culturing at 37 deg.C for 14 hr to obtain seed culture solution.
Yeast seed liquid: selecting YPD liquid culture medium (2% peptone, 1% yeast powder, 2% glucose), autoclaving at 115 deg.C for 20min, inoculating yeast at 2% inoculum size in YPD liquid culture medium, and culturing at 28 deg.C for 24 hr to obtain seed liquid.
Acetic acid bacteria seed liquid: selecting acetic acid bacteria liquid culture medium (1% yeast powder, 2% glucose), autoclaving at 115 deg.C for 20min, cooling, adding 3% anhydrous ethanol, inoculating acetic acid bacteria in liquid culture medium at 5% inoculum size, and culturing at 30 deg.C for 24 hr to obtain seed liquid.
(2) JYY015 strain was fermented, and the aroma components of the fermentation broth were measured and analyzed:
the yeast is cultured in YPD liquid medium for 36h, volatile components in the culture solution are collected by rotary evaporator, and aroma components are measured by gas chromatography-mass spectrometry (GC-MS). Gas Chromatography (GC) conditions: a flow splitting mode: no flow diversion; temperature rising procedure: the initial temperature is 40 ℃, the temperature is kept for 3min, then the temperature is increased to 120 ℃ at 3 ℃/min, and finally the temperature is increased to 230 ℃ at 8 ℃/min; carrier gas: helium (He); flow rate: 1 mL/min; detector temperature: 230 ℃, inlet temperature: 230 ℃ to 230 ℃.
Example 1, isolation and identification of aroma-producing bacteria and detection of aroma components:
(1) the separation of the aroma-producing yeast of the invention comprises the following steps: sampling fresh jujube juice from Ningxia Lingwu, returning to laboratory at low temperature, taking 10mL by aseptic technique, adding sterile physiological saline containing 90mL, shaking, performing gradient dilution in ultra-clean bench, and coating on YPD agar plate. The culture was inverted at 28 ℃ for 2 days for counting, 7 strains with typical yeast colony morphology were selected, numbered sequentially, inoculated and streaked onto YPD agar plates, and repeated 2-3 times until pure strains were obtained. Then inoculating 7 strains of bacteria into YPD liquid test tube, culturing at 28 deg.C for 2d, performing sensory evaluation on volatile flavor substances by olfaction sensitive personnel, wherein the fermentation liquid of No. 2 strain has strong fragrance, the strain name is modified to JYY015, and storing in-80 deg.C refrigerator.
(2) The JYY015 strain is subjected to microscopic examination, is in lemon type and bowling ball type (figure 1), is streaked on YPD solid culture medium, is cultured for 48h, has circular raised colony with regular edge, is milky white, has colony diameter of generally 3-5mm (figure 2), and is preliminarily judged to be a yeast from thallus size, smell and colony morphology.
(3) JYY015 single colony is picked and transferred into YPD liquid culture medium, and cultured for 24h, 2mL of fresh bacteria liquid is taken and sent to one generation for sequencing, and the detection sequence is 18S area rDNA. The detection result is entered into NCBI website BLAST, the comparison result is Hanseniaspora uvarum (Hanseniaspora uvarum), and the similarity reaches more than 99.9%.
(4) The grape juice Hansenula polymorpha JYY015 was deposited in the China general microbiological culture Collection center on 19.11.2019 at the location of Xilu No.1 Hospital, North Chen Yang district, Beijing. The preservation number is CGMCC No. 18969.
(5) The aroma-producing yeast is inoculated in YPD liquid culture medium with the inoculation amount of 2%, the YPD liquid culture medium is subjected to shaking culture at the temperature of 28 ℃ and the speed of 150rpm, the early growth is rapid, 8 hours enter a logarithmic phase, 15 hours enter a stationary phase, and the number of bacteria is reduced after 18 hours (figure 3).
(6) Aroma component analysis of aroma-producing bacteria JYY 015.
The GC-MS detects 102 volatile flavor substances which mainly comprise alcohol and ester substances. The material was screened for volatile content > 1% (see table 1). The content of the ethyl acetate is maximum, and the relative content reaches 29.8732 percent. The phenethyl acetate is sweet fragrance, has rose fragrance, has honey-like fragrance with powder fragrance, and also has fruit fragrance or peach fragrance. The second most relative content is phenethyl alcohol, and the relative content is 15.7874%. Has fresh bread fragrance and fresh and sweet rose fragrance. By analysis, JYY015 produced fragrance of high quality with overall floral notes biased towards floral, especially rose-like.
Example 2 application of the aroma-producing yeast JYY015 in Lingwu jujube ferment:
the application of the aroma-producing yeast in Lingwu jujube ferment comprises the following steps:
s1: pretreatment of raw materials: selecting Lingwu long jujube as raw material, cleaning, sterilizing, adding sterile water, breaking and homogenizing. Then filtering with 30 mesh filter sieve to remove residue, and pasteurizing the filtrate.
S2: adding auxiliary materials: adding 2% of sterilized sucrose, 10% of maltitol and 3% of inulin into the filtrate of the jujube juice. The disinfection method is ozone fumigation for 30 min. The auxiliary materials are required to be uniformly stirred in the adding process.
S3: and (3) bacteria planting: inoculating Hansenula sporulata JYY015, which is selected from the juice of Lingwu date and has local characteristics, to the selected grape juice, wherein the inoculation amount is 3%, the fermentation temperature is 28-30 ℃, and the fermentation time is 48-72 h.
S4: secondary bacterium planting: inoculating acetic acid bacteria and lactobacillus with inoculation amount of 5% for secondary fermentation at 30-33 deg.C, and controlling aeration fermentation in the fermentation tank for 24 hr to allow aerobic acetic acid bacteria to rapidly propagate and consume ethanol in the fermentation liquid. Anaerobic fermentation for 4-7 days to make lactobacillus become dominant strain, and pH is controlled at 3.2-3.6 after fermentation.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and improvements can be made without departing from the principle of the present invention, and the protection scope of the present invention is also considered.

Claims (4)

1. The yeast for producing the rose fragrance and the application thereof in Lingwu jujube ferment are characterized in that the yeast for producing the rose fragrance is Hansenula polymorpha JYY015 of grape juice with the preservation number of CGMCCNO.18969.
2. The rose-producing yeast according to claim 1, characterized in that the 18SrDNA sequence of the yeast is seq.id.no 1.
3. The yeast capable of producing rose flowers according to claim 1, wherein the yeast capable of producing rose flowers can be applied to Lingwu jujube ferment.
4. The use of the yeast with a floral rose aroma as claimed in claim 3 in Lingwu jujube ferment, characterized in that it comprises the following steps:
s1: pretreatment of raw materials: cleaning Lingwu long jujube, sterilizing, adding a certain proportion of sterile water, crushing and homogenizing, filtering with a 30-mesh filter sieve to remove residues, avoiding the influence of fruit residues on taste, and pasteurizing the filtrate.
S2: adding auxiliary materials: adding 2% of sterilized sucrose, 10% of sterilized maltitol and 3% of sterilized inulin into the filtrate of the jujube juice.
S3: and (3) bacteria planting: inoculating Hansenula polymorpha JYY015 to grape juice, fermenting at 28-30 deg.C for 48-72 h.
S4: secondary bacterium planting: inoculating acetic acid bacteria and lactobacillus, fermenting at 30-33 deg.C for 24 hr, sealing, and fermenting under anaerobic condition for 4-7 days with pH of 3.2-3.6.
CN202010396033.8A 2020-05-08 2020-05-08 Yeast capable of producing rose fragrance and application of yeast in Lingwu jujube enzyme Pending CN111607621A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112746029A (en) * 2021-01-22 2021-05-04 西北农林科技大学 Hansenula polymorpha strain QTX22 for producing aroma substances at high yield and application thereof
CN117721028A (en) * 2024-02-07 2024-03-19 天津科技大学 Hansenula polymorpha strain YN321 in grape juice and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
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