CN117866787A - Aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof - Google Patents
Aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof Download PDFInfo
- Publication number
- CN117866787A CN117866787A CN202410096924.XA CN202410096924A CN117866787A CN 117866787 A CN117866787 A CN 117866787A CN 202410096924 A CN202410096924 A CN 202410096924A CN 117866787 A CN117866787 A CN 117866787A
- Authority
- CN
- China
- Prior art keywords
- aroma
- yeast
- content
- phagostimulant
- strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 74
- 239000000796 flavoring agent Substances 0.000 title claims abstract description 20
- 235000019634 flavors Nutrition 0.000 title claims abstract description 19
- 235000013399 edible fruits Nutrition 0.000 title claims abstract description 11
- WRMNZCZEMHIOCP-UHFFFAOYSA-N 2-phenylethanol Chemical compound OCCC1=CC=CC=C1 WRMNZCZEMHIOCP-UHFFFAOYSA-N 0.000 claims abstract description 36
- 238000000855 fermentation Methods 0.000 claims abstract description 35
- 230000004151 fermentation Effects 0.000 claims abstract description 35
- 239000000126 substance Substances 0.000 claims abstract description 18
- 241000178263 Dipodascus macrosporus Species 0.000 claims abstract description 14
- 230000012010 growth Effects 0.000 claims abstract description 11
- 241000220317 Rosa Species 0.000 claims abstract description 5
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 230000001580 bacterial effect Effects 0.000 claims description 17
- 239000003205 fragrance Substances 0.000 claims description 16
- 239000008369 fruit flavor Substances 0.000 claims description 14
- 108020004463 18S ribosomal RNA Proteins 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims 1
- 239000007788 liquid Substances 0.000 abstract description 14
- 238000012216 screening Methods 0.000 abstract description 6
- 230000002045 lasting effect Effects 0.000 abstract description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 58
- 239000001963 growth medium Substances 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 7
- 244000168141 Geotrichum candidum Species 0.000 description 6
- 235000017388 Geotrichum candidum Nutrition 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 241000220225 Malus Species 0.000 description 5
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 238000004451 qualitative analysis Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000012764 semi-quantitative analysis Methods 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- 241000235645 Pichia kudriavzevii Species 0.000 description 4
- 125000003118 aryl group Chemical group 0.000 description 4
- 150000002148 esters Chemical class 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 235000019629 palatability Nutrition 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000007222 ypd medium Substances 0.000 description 4
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241000235648 Pichia Species 0.000 description 3
- 239000003674 animal food additive Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000011218 seed culture Methods 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 241000228212 Aspergillus Species 0.000 description 2
- 206010011732 Cyst Diseases 0.000 description 2
- 241000235036 Debaryomyces hansenii Species 0.000 description 2
- 244000286779 Hansenula anomala Species 0.000 description 2
- 235000014683 Hansenula anomala Nutrition 0.000 description 2
- 241000235644 Issatchenkia Species 0.000 description 2
- 241001123674 Metschnikowia Species 0.000 description 2
- 241001489192 Pichia kluyveri Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 230000002421 anti-septic effect Effects 0.000 description 2
- 235000021016 apples Nutrition 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 208000031513 cyst Diseases 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- XIRNKXNNONJFQO-UHFFFAOYSA-N ethyl hexadecanoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC XIRNKXNNONJFQO-UHFFFAOYSA-N 0.000 description 2
- MMXKVMNBHPAILY-UHFFFAOYSA-N ethyl laurate Chemical compound CCCCCCCCCCCC(=O)OCC MMXKVMNBHPAILY-UHFFFAOYSA-N 0.000 description 2
- RRAFCDWBNXTKKO-UHFFFAOYSA-N eugenol Chemical compound COC1=CC(CC=C)=CC=C1O RRAFCDWBNXTKKO-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- WOFPPJOZXUTRAU-UHFFFAOYSA-N octan-4-ol Chemical compound CCCCC(O)CCC WOFPPJOZXUTRAU-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000001953 sensory effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 235000015096 spirit Nutrition 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001465178 Bipolaris Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- NLZUEZXRPGMBCV-UHFFFAOYSA-N Butylhydroxytoluene Chemical compound CC1=CC(C(C)(C)C)=C(O)C(C(C)(C)C)=C1 NLZUEZXRPGMBCV-UHFFFAOYSA-N 0.000 description 1
- NPBVQXIMTZKSBA-UHFFFAOYSA-N Chavibetol Natural products COC1=CC=C(CC=C)C=C1O NPBVQXIMTZKSBA-UHFFFAOYSA-N 0.000 description 1
- 235000009917 Crataegus X brevipes Nutrition 0.000 description 1
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 description 1
- 235000009685 Crataegus X maligna Nutrition 0.000 description 1
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 description 1
- 235000009486 Crataegus bullatus Nutrition 0.000 description 1
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 description 1
- 235000009682 Crataegus limnophila Nutrition 0.000 description 1
- 240000000171 Crataegus monogyna Species 0.000 description 1
- 235000004423 Crataegus monogyna Nutrition 0.000 description 1
- 235000002313 Crataegus paludosa Nutrition 0.000 description 1
- 235000007106 Crataegus suborbiculata Nutrition 0.000 description 1
- 241000073432 Crataegus suborbiculata Species 0.000 description 1
- 235000009840 Crataegus x incaedua Nutrition 0.000 description 1
- 241001123635 Dipodascus Species 0.000 description 1
- 239000005770 Eugenol Substances 0.000 description 1
- 240000002045 Guettarda speciosa Species 0.000 description 1
- 235000001287 Guettarda speciosa Nutrition 0.000 description 1
- 241001149669 Hanseniaspora Species 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- 241001123676 Metschnikowia pulcherrima Species 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- UVMRYBDEERADNV-UHFFFAOYSA-N Pseudoeugenol Natural products COC1=CC(C(C)=C)=CC=C1O UVMRYBDEERADNV-UHFFFAOYSA-N 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 241000235029 Zygosaccharomyces bailii Species 0.000 description 1
- 235000013202 a hawthorn Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- -1 aldehyde ketones Chemical class 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 239000005667 attractant Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 230000031902 chemoattractant activity Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000011363 dried mixture Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940067592 ethyl palmitate Drugs 0.000 description 1
- 229960002217 eugenol Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 235000019990 fruit wine Nutrition 0.000 description 1
- 235000021021 grapes Nutrition 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000010813 internal standard method Methods 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000033458 reproduction Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000010563 solid-state fermentation Methods 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
- 235000021419 vinegar Nutrition 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Abstract
The invention relates to the technical field of bioengineering, in particular to aroma-producing yeast for fermenting and presenting fruit aroma flavor and application thereof, and the strain belongs to Geopodiopsis grossedentata Dipodascus macrosporus DDC008 with a preservation number of CCTCC M20232692. Separating and screening mature apple peel from Fufeng county in Shaanxi to obtain a strain of aroma-producing yeast Geopodiopsis glabra (Dipodascus macrosporus), wherein the optimal growth condition of the strain is that the temperature is 28 ℃ and the pH is 4.0; the aroma-producing yeast synthesizes the beta-phenethyl alcohol in the liquid fermentation process, the substance has soft, pleasant and lasting rose aroma, the content of the beta-phenethyl alcohol accounts for 60 percent or more of the total content of volatile substances, the shake flask level semi-quantitatively determines the content of 100mg/L, and the pilot test level semi-quantitatively determines the content of 1g/L.
Description
Technical Field
The invention relates to the technical field of bioengineering, in particular to aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof.
Background
The aroma-producing yeasts are called aroma-producing yeasts and ester-producing yeasts, belong to non-saccharomyces cerevisiae and refer to all yeasts except saccharomyces cerevisiae strains, and are yeasts capable of producing aromatic substances in the growth and metabolism process, wherein the aromatic substances are mainly esters, and volatile flavor substances such as alcohols, acids, aldehydes, ketones, phenols, terpenes and the like. Mainly comprises Pichia (Pichia), candida, hansenula (Hanseniaspora), issatchenkia (Issatchenkia), metschnikowia (Metschnikowia) and the like, wherein representative yeast strains are Pichia kluyveri (Pichia kluyveri), torulopsis variabilis (Torulopsis versatilis), debaryomyces hansenii (Debaryomyces hansenii), issatchenkia orientalis (Issatchenkia orientalis), megazelle (Metschnikowia pulcherrima) and the like.
The aroma-producing yeast has wide application fields, and is widely applied and researched in the fields of white spirit, grape wine, table vinegar, fruit wine, fruit juice, soy sauce, pickled vegetable and other fermented foods, tobacco fermentation, biological fermentation feed, microbial synthesis essence and spice and the like. Especially in the white spirit brewing industry, flavor substances in various aromatic white spirits are related to various genus aroma-producing yeasts such as Pichia anomala (Pichia anomala), pichia kudriavzevii (P.kudriavzevii), boehringer's joint yeast (Zygosaccharomyces bailii) and the like in fermented grains of white spirits or Daqu. At present, the screening and pure culture of the aroma-producing yeast are realized, and the aroma-producing yeast or aroma-producing microorganism has wide application prospect in the future.
Although it is not yet known whether the asexual stage belongs to Geotrichum candidum (Geotrichum candidum) as an aromatic microorganism between yeast and fungi, geotrichum candidum (Dipodascus macrosporus) is one of important yeast groups, and a large number of fungi of the genus Dipodascus have been found by Xu Yan research team to appear in fermented grains of Luzhou-flavor liquor, which has an important effect on the flavor of the liquor. In recent years, research and application of a related microorganism Geotrichum candidum (Geotrichum candidum) of Geotrichum candidum (Dipodascus macrosporus) and fermentation products thereof are attracting attention, and the method is widely applied to food industry, brewing industry, medicine industry, feed industry and the like. The research report about Geopodiopsis grossedentata (Dipodascus macrosporus) is sporadic, the basic and application researches are not deep, and the related report about the fermentation aroma-producing performance and application of the pure yeast strain is not found. The invention applies the aroma-producing microorganism of the Geopodiopsis grossedentata (Dipodascus macrosporus) and a fermentation product thereof to the field of feed additives, and adopts a microorganism fermentation mode to prepare the feeding attractant. Compared with the traditional feed additive, the fermentation phagostimulant has the advantages of more abundant nutrient substances, improved palatability and more natural and mellow fragrance, and provides a new scene of biological application of the bipedal cyst fungus (Dipodascus macrosporus).
Disclosure of Invention
The invention aims to provide aroma-producing yeast capable of fermenting to form fruit aroma and application thereof, and solves the technical problems of single aroma, insufficient flavor, mellow taste, poor palatability and the like of a traditional feed additive in the prior art.
The invention discloses aroma-producing yeast with fruit aroma flavor by fermentation, which belongs to Geopodiopsis grossedentata Dipodascus macrosporus DDC with a preservation number of CCTCC M20232692.
The strain has been preserved in China Center for Type Culture Collection (CCTCC) at a preservation address of university of Wuhan, china, 12 months and 27 days.
Wherein the yeast with fruit flavor is obtained by separating pericarp of mature apple in Fufeng county in Shanxi province.
Wherein, the yeast with fruit flavor by fermentation has the following growth conditions: the temperature is 25-32 ℃, and the pH is 3.0-8.0.
Preferably, the yeast growth conditions for the fermentation to have fruit flavor are as follows: the temperature was 28℃and the pH4.0.
Wherein, the high-yield beta-phenethyl alcohol of the yeast which is fermented to be fruity flavor has soft and durable rose fragrance, the content of the beta-phenethyl alcohol accounts for 50-80 percent of the total content of volatile substances, and the semi-quantitative determination content is 100mg/L;
the biological characteristics of the yeast fermented to have fruit flavor are as follows: after 24 hours of cultivation on YPD medium at 28 ℃, white, irregular and fluffy moist micro-convex colonies are formed, the characteristic fragrance of apples is obvious, the cells are nearly rectangular, and budding and reproduction are not seen.
Wherein the composition of the YPD medium comprises: 20g of glucose, 10g of yeast powder, 20g of peptone and 10g of agar in every 1000mL of water. Sterilizing at 115deg.C under high pressure for 15min.
Wherein, the yeast which is fermented to have fruit flavor is fermented for 30 hours in a 50L stirring type bioreactor to obtain the bacterial cells with the wet weight of 200g/L, and the content of the beta-phenethyl alcohol is detected for 1g/L in a semi-quantitative way for 48 hours.
The application of aroma-producing yeast with fruit aroma flavor by fermentation is used for preparing phagostimulant.
Further, the phagostimulant is a feeding phagostimulant or an edible phagostimulant.
A feeding phagostimulant is prepared from the aroma-producing yeast strain.
Further, the specific steps are that the bacterial liquid with the wet weight content of the aroma-producing yeast strain being more than 20g/100g, the adsorption carrier and the food-grade antiseptic bacteriostat are mixed and adsorbed, and the aroma-producing yeast strain is obtained after drying.
Further, the bacterial liquid is bacterial cells and fermentation liquid.
Further, the drying temperature is 65-75 ℃.
Further, the drying controls the moisture content to be within 10%.
An edible phagostimulant is prepared from the above aroma-producing yeast strain.
Compared with the prior art, the invention has the following beneficial effects:
1. separating and screening mature apple peel from Fufeng county in Shaanxi to obtain a strain of aroma-producing yeast Geopodiopsis glabra (Dipodascus macrosporus), wherein the optimal growth condition of the strain is that the temperature is 28 ℃ and the pH is 4.0;
2. the aroma-producing yeast synthesizes beta-phenethyl alcohol in the shake flask horizontal liquid state fermentation process, the substance has soft, pleasant and lasting rose aroma, the content of the beta-phenethyl alcohol accounts for 60 percent or more of the total content of volatile substances, and the semi-quantitative determination content is 100mg/L;
3. the aroma-producing yeast is cultured for 30 hours in a 50L stirring type bioreactor at high density, and finally the thallus cells with the wet weight of 200g/L are obtained; semi-quantitatively detecting the content of the beta-phenethyl alcohol by 1g/L for 48 hours;
4. the animal phagostimulant of the invention presents sweet smell, thick fermentation aroma and fruit aroma taste of fermented glutinous rice, is detected in a sterile way, and is placed at normal temperature to keep aroma for 6 months or more; the yeast disclosed by the invention improves the flavor of the fermented feeding phagostimulant, has mellow and mild fruit flavor characteristics, is better in palatability and improves the total volatile matters.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a diagram showing the morphology and colony morphology of Dipodascus macrosporus of the present invention.
FIG. 2 shows a growth curve of the invention Dipodascus macrosporus.
FIG. 3 is a diagram showing the upper tank culture of Dipodascus macrosporus of the present invention.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments.
Example 1
The embodiment discloses a aroma-producing yeast fermented to have fruit aroma flavor, and the screening method comprises the following steps:
01 Primary separation and purification of aroma-producing yeast
Sampling from mature apples produced in Fufeng county of Shaanxi, harrow oranges in Zhongjiang county of Sichuan and local grapes of Chengdu, taking pulp and pericarp respectively, placing into a sterile conical flask, culturing for 3d in a culture box at 28 ℃, and adding 50ml of sterile water to prepare suspension. The bacterial suspension is used as mother solution to be diluted to 10 in gradient -6 Take 10 -4 -10 -6 The diluted bacterial liquid is coated on an enrichment culture medium, inverted culture is carried out for 48 hours at 28 ℃, single bacterial colonies with different bacterial colony forms are picked for microscopic examination and observation, strains with suspected yeast single cell forms are transferred to the liquid culture medium one by one, and numbering and preservation are carried out; after the cultivation is finished, a proper amount of bacterial liquid is taken, sensory evaluation is carried out by a sniffing method, whether aroma is generated or not is carried out, aroma characteristics and intensity are distinguished, more than ten aroma-producing yeast strains are screened out, and 3 strains are sensory evaluated to be good in aroma-producing performance.
02 Screening of aroma-producing yeasts
Inoculating 3 strains with excellent aroma producing performance obtained by separation and purification into a yeast extract peptone glucose culture medium, culturing for 48 hours, accurately weighing 1.5g of bacterial liquid, treating, and carrying out qualitative and semi-quantitative analysis by using an internal standard method through solid phase microextraction-gas chromatography mass spectrometry to obtain the strain DDC008 with the best aroma producing performance.
Example 2
The screened DDC008 strain is subjected to physiological and biochemical characteristic identification according to microbial taxonomy (see tables 1 and 2), genomic DNA is extracted according to a yeast genome extraction kit extraction method, and P1 and P2 are used as forward and reverse primers:
P1：5'-GTAGTCATATGCTTGTCTC-3'，
P2：5'-GCATCACAGACCTGTTATTGCCTC-3'。
the 18S rDNA gene was amplified by PCR, and 18S rDNA sequencing was carried out by the company of Shanghai, inc., to obtain the 18S rDNA sequence of the strain, and then homology alignment was carried out on NCBI website by BLAST search tool. Based on 18S rDNA sequence analysis and physiological and biochemical characteristic identification, the strain is considered to be bipedal cyst strain and named Dipodascus macrosporus DDC008 in combination with research on the growth and fermentation characteristics of the strain.
SEQ ID NO. 1 Strain DDC008 part 18S rDNA sequence:
1TAACTACTCC CCCAGACCCA AAAACTTTGATTTCTCGTAAGGTGCCGGAT GGGTTTATGT
61 AACGCCATCC GATCCCTAGT CGGCATAGTTTACGGTTAAG ACTACGACGG TATCTGATCG
121 TCTTCGATCC CCTAACTTTC GTTCTTGATTAATGAAAACG TCCTTGGCAA ATGCTTTCGC
181 AGTAGTTAGT CTTCAGTAAA TCCAAGAATTTCACCTCTGA CAACTGAATA CTGATGCCCC
241 CGACCGTCCC TATTAATCAT TACGACGGTCCTAGAAACCA ACAAAATAGAACCATACGTC
301 CTATTCTATT ATTCCATGCT AATATATTCGAGCAAAGGCC TGCTTTGAAC ACTCTAATTT
361 TTTCAAAGTAAATGTTTGGT TTATTCTCCCTAAATAAATAGAGAGCTTCC ACCAAAGAAA
421 GATGTTTTGT GGAAGTACTT AAAAAAAGGCCCCCCCGTCA ACATCCAAGT TTCAACTACG
481 AGCTTTTTAA CTGCAACAAC TTTAATATATACTATCAGAG CTGGAATTAC CGCGGCTGCT
541 GGCACCAGAC TTGCCCTCTA ATTGTTCCTCGTTAAGGTAT TTAAATTGTT CTCATTCCAA
601 TTACGAGACC TAATAGGCCC CGTATCGTTATTTATTGTCA CTACCTCCCT GTGTCAGGAT
661 TGGGTAATTT GCGCGCCTGC TGCCTTCCTTGGATGTGGTA GCCGTTTCTC AGGCTCCCTC
721 TCCGGAATCG AACCCTGATT CCCCGTTACC CGTTAAAACC ATGGTAGGCC TCTATCCTAC
781 CATCGAAAGT TGATAGGGCA GAAATTTGAA TGAACCATCG CCGGCACAAG GCCATGCGAT
841 TCGACAAGTT ATTATGATTC ACCATAGAGC CCGAAGGCAT TGGTTTTTTA TCTAATAAAT
901 ACCAGCCGGT AAACCGGCCG TTTTAGCATG TATTAGCTCT AGAATTACCA CGGTTATCCA
961 TGTAGTAATG TAATATCAAA TAAACGATAA CTGATTTAAT GAGCCATTCG CAGTTTCACT
1021 GTATAATGCT TATACTAGAC ATCAGG
TABLE 1 physiological and biochemical identification results of Strain (DDC 008)
TABLE 2 fragrance intensity and characteristics of Strain (DDC 008) fermented in YPD medium for 24h and 48h
And (3) injection: the "++ + +" indicates that the fragrance is intense and the fragrance pleasure degree is high; "++" indicates slightly lighter fragrance and less pleasant fragrance; "+" indicates that the fragrance is slight and the pleasure of the fragrance is slight.
TABLE 3 fragrance substance case where Strain (DDC 008) was fermented in YPD medium for 48h
Example 3
Step 1: preparation of the culture Medium
Seed medium (g/L): glucose 20, peptone 20, yeast powder 10, initial pH 5.0-5.5;
fermentation medium (g/L): glucose 20, peptone 20, yeast powder 10, initial pH 5.0-5.5;
step 2: seed preparation
Seed medium in 500mL Erlenmeyer flask was 50mL and sterilized at 115℃for 15min. The strain is preserved in an glycerol pipe with the final concentration of 15%, 200 mu L of preserved bacterial liquid is inoculated into 50ml of seed culture medium for seed culture, and the culture is carried out at 28 ℃ for 12-14h at 200 rpm.
Step 3: shaking flask fermentation medium
The seed culture medium is inoculated with 200mL of fermentation culture medium for fermentation culture with 10% of inoculation amount. Fermenting at 28deg.C and 200rpm for 48 hr, accurately weighing 1.5g of fermentation broth, processing, adding internal standard 4-octanol, and performing qualitative and semi-quantitative analysis of volatile substances by solid phase microextraction-gas chromatography-mass spectrometry. Phenethyl alcohol 100mg/L.
Example 4
Fermentation culture in 50L fermentation tank:
step 1: same as in example 3
Step 2: same as in example 3
Step 3: fermentation culture in 50L fermentation tank
The liquid loading amount of a fermentation medium in a 50L fermentation tank is 30L, the inoculation amount is 10% (v/v), the temperature is 28 ℃, the stirring speed is 500r/min, the ventilation amount is 30L/min, the pH is controlled in the logarithmic phase of growth, the pH is not controlled in the platform phase and later, and the culture time is 40-48h. Analyzing the wet weight of the cells; 1.5g of zymophyte liquid is accurately weighed for treatment, an internal standard 4-octanol is added, and qualitative and semi-quantitative analysis of volatile substances is carried out by adopting solid-phase microextraction-gas chromatography-mass spectrometry. The wet weight of the bacterial cells is 20g/100g; 1g/L of phenethyl alcohol.
As shown in fig. 1-3 and in tables 1-3, a aroma-producing yeast is obtained by separating and screening mature apple peel from Fufeng county of Shaanxi, and the bacterial colony is white, round, dry and opaque, has fluff at irregular edges, is not smooth and is tightly wetted and tiled, and has typical yeast aroma; the cell morphology is rectangular, obvious spores are not seen, and the strain is identified as the bipolaris geodesica. The optimal growth temperature of the strain is 28 ℃, the pH value is 4.0, the beta-phenethyl alcohol is synthesized in the liquid fermentation process in YPD culture medium, the substance has soft, pleasant and lasting rose fragrance, the content of the beta-phenethyl alcohol accounts for 60% or more of the total content of volatile substances, and the semi-quantitative determination content is 100mg/L. Culturing for 30h in a 50L stirring type bioreactor at high density to finally obtain the bacterial cells with the wet weight of 200g/L, and semi-quantitatively determining the content of the beta-phenethyl alcohol for 48h to be 1g/L.
Example 5
Animal phagostimulant prepared by using yeast of the invention
Step 1: same as in example 4
Step 2: same as in example 4
Step 3: same as in example 4
Step 4: animal phagostimulant formulation
According to the characteristic of the invention, the yeast is fermented to synthesize the flavor substance, the bacterial liquid (bacterial cells and fermentation liquid) with the wet weight content of more than 20g/100g, the adsorption carrier and the food-grade antiseptic bacteriostat are mixed and adsorbed, and the mixture is dried at the high temperature of 65-75 ℃ with the moisture content controlled within 10%, and the dried mixture is the animal phagostimulant. The product has sweet smell, strong fermentation aroma and fruit aroma, no bacteria detection, and fragrance retention at normal temperature for 6 months or more. GC-MS qualitative and semi-quantitative analysis shows that the volatile matters of the animal phagostimulant prepared by the yeast comprise about 32% of alcohols, about 22% of esters, about 14% of acids and about 25% of aldehyde ketones, and the phagostimulant product also comprises active matters such as proline, antioxidant KB and the like. The animal phagostimulant can be used for animals such as fish, pig, cattle and sheep.
Example 6
The yeast of the invention is used for improving the flavor of the fermented feeding phagostimulant:
step 1: same as in example 3
Step 2: same as in example 3
Step 3: solid state fermentation culture
According to the characteristics of quick growth and good flavor of the yeast, the yeast is inoculated into a feeding phagostimulant for fermentation. The logarithmic phase yeast seed solution is obtained according to example 3, inoculated into a feeding phagostimulant taking hawthorn powder as a main raw material according to an inoculum size of 10%, mixed with commercial Angelica yeast (aspergillus, saccharomyces cerevisiae) for fermentation for 5-7 days, dried at a high temperature of 70 ℃ and the moisture content is controlled within 10%, and crushed into powder. Compared with single commercial Angel yeast (aspergillus, saccharomyces cerevisiae) phagostimulant, the yeast participated fermentation phagostimulant of the invention not only maintains the characteristic fruit flavor of a hawthorn powder fermentation substrate, but also presents fresh and soft flower and fruit flavor, and has more mellow fruit flavor and better palatability, and the GC-MS qualitative and semi-quantitative analysis shows that the total improvement of volatile substances is about 25.6%, the esters account for more than 85%, wherein the phenethyl alcohol content is improved by about 42.3%, the eugenol content is improved by about 68.5%, the ethyl laurate content is improved by about 26.7%, the ethyl palmitate content is improved by about 70.8%, and the like.
The above is an embodiment exemplified in this example, but this example is not limited to the above-described alternative embodiments, and a person skilled in the art may obtain various other embodiments by any combination of the above-described embodiments, and any person may obtain various other embodiments in the light of this example. The above detailed description should not be construed as limiting the scope of the present embodiments, which is defined in the claims and the description may be used to interpret the claims.
Claims (10)
1. A aroma-producing yeast which is fermented to have fruit aroma flavor, which is characterized in that: the strain belongs to Geopodiopsis grossedentata Dipodascus macrosporus DDC008 and has a preservation number of CCTCC M20232692.
2. The yeast fermented to fruit flavor according to claim 1, wherein: the content of beta-phenethyl alcohol produced by the yeast accounts for 50-80% or more of the total content of volatile substances, and the beta-phenethyl alcohol has rose fragrance.
3. The yeast fermented to fruit flavor according to claim 1, wherein: the semi-quantitative determination content of the beta-phenethyl alcohol is 100mg/L.
4. The yeast fermented to fruit flavor according to claim 1, wherein: the growth conditions are at a temperature of 25-32deg.C, preferably 28deg.C, and pH3.0-8.0, preferably pH4.0.
5. The yeast fermented to fruit flavor according to claim 1, wherein: the 18S rDNA sequence of the yeast is shown as SEQ ID NO. 1.
6. The yeast fermented to fruit flavor according to claim 1, wherein: the inoculation amount is 10%, fermentation culture is carried out for 30 hours in a 50L stirring type bioreactor, the wet weight of the bacterial cells is 200g/L, and the content of the beta-phenethyl alcohol is 1g/L in 48 hours semi-quantitative measurement.
7. Use of a aroma-producing yeast that ferments to a fruity flavour according to any one of claims 1-6, characterized in that: is used for preparing phagostimulant.
8. The use of a aroma-producing yeast that ferments to a fruity flavor according to claim 7, characterized in that: the phagostimulant is a feeding phagostimulant or an edible phagostimulant.
9. A feeding phagostimulant, which is characterized in that: use of a aroma-producing yeast strain according to claim 1.
10. An edible phagostimulant, which is characterized in that: use of a aroma-producing yeast strain according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410096924.XA CN117866787A (en) | 2024-01-23 | 2024-01-23 | Aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410096924.XA CN117866787A (en) | 2024-01-23 | 2024-01-23 | Aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117866787A true CN117866787A (en) | 2024-04-12 |
Family
ID=90592985
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410096924.XA Pending CN117866787A (en) | 2024-01-23 | 2024-01-23 | Aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117866787A (en) |
-
2024
- 2024-01-23 CN CN202410096924.XA patent/CN117866787A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104397632B (en) | Fermented capsicum product and preparation method thereof | |
| JP7417964B2 (en) | Saccharopolyspora and its use in reducing biogenic amines | |
| CN111961615B (en) | Saccharopolyspora capable of reducing biogenic amine and application thereof | |
| CN109679855B (en) | Eurotium shiatu producing tea-flavor and application thereof in white spirit yeast | |
| KR101082246B1 (en) | Nuruk containing salicornia herbacea and preparation method of the same | |
| CN113234639B (en) | Lactobacillus plantarum ZF632 and application thereof | |
| CN102352323A (en) | Ester producing yeast as well as method and application of yeast for producing Xiaoqu fen-flavor seasoning wine | |
| CN105018384A (en) | Acetobacter strain and application thereof in fermenting apple vinegar | |
| CN105767984A (en) | Fermented chili-pepper product and preparation method thereof | |
| CN111979148B (en) | Saccharopolyspora composition and application thereof in food | |
| CN111248409A (en) | Low-salt thick broad-bean sauce fermentation method | |
| CN105942299A (en) | Method for preparing functional minced larimichthys crocea products by using mixed microbe fermentation | |
| CN114591850A (en) | Saccharomyces cerevisiae CMRC 5S and application thereof | |
| CN113249268B (en) | Saccharopolyspora rosea for reducing biogenic amine and application thereof | |
| CN112592857B (en) | Microbial agent for fermentation of aged daocai | |
| CN111635870B (en) | Leavening agent, direct-feeding fermentation method of stinky tofu marinating liquid and stinky tofu marinating liquid | |
| CN113151023A (en) | Aroma-enhancing type direct-vat-set starter, and preparation method and application thereof | |
| CN116376729A (en) | Wick yeast, microbial preparation and medlar western style wine and brewing method thereof | |
| CN117229929A (en) | High-yield ester abnormal Wicks yeast and application thereof | |
| CN117866787A (en) | Aroma-producing yeast capable of fermenting to form fruit aroma flavor and application thereof | |
| CN107916194A (en) | A kind of method that elaeagnus conferta fruits fermented wine is prepared with brown sugar and elaeagnus conferta fruits | |
| JP5478692B1 (en) | Novel fermented flavor liquid and food containing the fermented flavor liquid | |
| CN113265363B (en) | Saccharopolyspora cholerae for reducing biogenic amine and application thereof | |
| JP7329221B2 (en) | Saccharopolyspora composition and its use in food | |
| Prasad et al. | Fate of acidity and alcohol during fermentation of Salalah and Sri Lanka coconut water |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination |