CN117562931A

CN117562931A - Composition containing Kluyveromyces marxianus thallus, preparation method and application

Info

Publication number: CN117562931A
Application number: CN202410051597.6A
Authority: CN
Inventors: 骆滨; 王菁; 吴蓉; 许昱; 牛志华; 张猛; 杨秀华; 邱博书; 葛凯
Original assignee: Shanghai Changjin Biotechnology Co ltd
Current assignee: Shanghai Changjin Biotechnology Co ltd
Priority date: 2024-01-15
Filing date: 2024-01-15
Publication date: 2024-02-20
Anticipated expiration: 2044-01-15

Abstract

The disclosure relates to the field of biotechnology, and in particular relates to a composition containing kluyveromyces marxianus thallus, a preparation method and application thereof. The inventor tries to use kluyveromyces marxianus or a composition containing kluyveromyces marxianus in preparation of medicines and/or foods, and verifies that various compositions containing different kluyveromyces marxianus strain thalli provided by the disclosure can remarkably improve compliance of users, and has development prospects of microbial medicines or functional foods.

Description

Composition containing Kluyveromyces marxianus thallus, preparation method and application

Technical Field

The disclosure relates to the field of biotechnology, and in particular relates to a composition containing kluyveromyces marxianus thallus, a preparation method and application thereof.

Background

Yeast is widely used in traditional and modern biotechnology, mainly for the production of pharmaceuticals, foods, beverages, enzymes, fine chemicals, etc., where the use of Saccharomyces cerevisiae and its related "conventional" yeasts in fermented beverages is well known.

With the great development of modern molecular biology, the potential value of some so-called "unconventional" yeasts is fully exploited, becoming increasingly important and becoming a research hotspot, including kluyveromyces marxianus.

Kluyveromyces marxianus (Kluyveromyces Marxianus) is a semi-fungal yeast, has close relation with Saccharomyces cerevisiae in genetics, is sister species with Kluyveromyces lactis, and can absorb lactose as a sugar source, and is a remarkable characteristic of the Kluyveromyces marxianus and Kluyveromyces lactis different from the traditional yeasts. This feature allows kluyveromyces marxianus and kluyveromyces lactis to be easily separated from dairy products, such as fermented milk, yogurt, cheese, and the like, and therefore, kluyveromyces marxianus and kluyveromyces marxianus quickly pass the GRAS and QPS security certificates of the united states and the european union.

Kluyveromyces lactis has been the main research species of kluyveromyces lactis, originally used for lactose metabolism research and later used as a model of unconventional yeasts, but as research proceeds, it was found that some of the characteristics of kluyveromyces marxianus are more suitable for the requirements of industrialization, such as extremely fast growth rate, heat resistance and high secretion capacity.

The industrial application of kluyveromyces marxianus is mainly used for producing natural enzymes, lactonase, beta-galactosidase, pectinase and the like, and the expression capacity of the enzymes enables the kluyveromyces marxianus to be used for treating lactose-containing waste in cheese industry or used for producing biomass or bioethanol. At present, kluyveromyces marxianus is commonly used for preparing a fermentation product with edible value through in-vitro fermentation, and also relates to preparation of a fermentation product with medicinal value. However, in the prior report, the fermentation product is obtained by in vitro fermentation and then directly eaten to play the edible or medicinal functions, whether the edible product or the medicinal product is prepared. However, the odor of the bacterial cells significantly reduces the medical compliance and food taste of the user, and is not conducive to popularization and application, and therefore, the taste masking of kluyveromyces marxianus for pharmaceutical or food applications is required.

Currently, taste masking agents used include beta-cyclodextrin, flavanones, magnesium sulfate, fatty acids, etc., but these are often chemical additives, have no medicinal or nutritional value per se, even have side effects, or have unsatisfactory mouthfeel. Some taste masking agents hinder the exertion of the medicinal and food functions of kluyveromyces marxianus per se and lose the medicinal effects, and therefore, there is still a need to develop novel taste masking agents or pharmaceutical or food compositions containing kluyveromyces marxianus which can be used for pharmaceutical or food compositions containing kluyveromyces marxianus or which are satisfactory in taste.

Disclosure of Invention

The present disclosure aims to provide a composition containing kluyveromyces marxianus, wherein the kluyveromyces marxianus and intracellular metabolites thereof selected in the composition have medicinal value, and the reasonable compounding of the composition is expected to be utilized to change or eliminate the peculiar smell of the kluyveromyces marxianus.

In order to solve the above technical problems and achieve the above objects, the present disclosure provides the following technical solutions:

in a first aspect, the present disclosure provides a composition comprising kluyveromyces marxianus, the composition comprising at least two components having a characteristic recognition odor, and the composition after the components having the characteristic recognition odor are compounded with kluyveromyces marxianus having a characteristic recognition odor different from any of the components in the composition.

In an alternative embodiment, the Kluyveromyces marxianus comprises at least one of a preservation number of CCTCC No. M20211265, CCTCC No. M20211601, CCTCC No. M20211602, CCTCC No. M20211603, CCTCC No. M20211604, CCTCC No. M20211605, CCTCC No. M20211606, CCTCC No. M20211607, CCTCC No. M20211608, CCTCC No. M20211609 or CCTCC No. M20211610.

In alternative embodiments, the component having a typical identifying odor comprises a nutty component, and/or a spicy Xin Liaowei component.

In alternative embodiments, the nut-flavored component comprises a cashew nut-flavored component, a walnut-flavored component, a chestnut-flavored component, a pistachio-flavored component, a hawaii-flavored component, an almond-flavored component, a badam-flavored component, a pine-flavored component, a gingko-flavored component, a lotus seed-flavored component, a peanut-flavored component, a chickpea-flavored component, or a sunflower-flavored component; preferably cashew nut taste components.

The spicy Xin Liaowei component comprises a garlic component, a green onion component or an onion component, preferably a garlic component.

In an alternative embodiment, the composition comprises, by mass, 10-20 parts of kluyveromyces marxianus thallus powder, 74.4-84.7 parts of nut taste components and 0.8-1.2 parts of spicy Xin Liaowei components. Wherein the nut-flavored component is preferably cashew dust, the cashew dust comprises a fried cashew dust comprising palm-oil-fried cashew dust and/or coconut-oil-fried cashew dust; the spicy Xin Liaowei component is preferably garlic powder.

In an alternative embodiment, the composition further comprises a salt.

In alternative embodiments, the palm oil is selected from 58 degrees palm oil, 52 degrees palm oil, 44 degrees palm oil, 33 degrees palm oil, or 24 degrees palm oil; the coconut oil is selected from purified coconut oil or cold pressed coconut oil. The frying temperature of the fried cashew nut powder is 120-150 ℃, and the frying time is 1-5 min.

In alternative embodiments, the composition further comprises a powder flavor and/or a liquid flavor; according to mass fraction, the powder essence is 0.2-0.3 part, and the liquid essence is 0.1-0.15 part; the aroma of the powder essence and the liquid essence is selected from cheese flavor or cheese flavor.

In a third aspect, the present disclosure provides a method of preparing a composition according to any one of the preceding embodiments, the method comprising mixing a component having a typical recognition odor with kluyveromyces marxianus thallus powder and crushing into a powder.

In a fourth aspect, the present disclosure provides the use of kluyveromyces marxianus, the composition of any one of the foregoing embodiments, or a composition obtained by the method of preparation of the foregoing embodiments, in the preparation of a microbial agent; the microbial agent is used for preventing allergy, assisting in treating helicobacter pylori, reducing cholesterol, relieving diarrhea and enteritis, regulating intestinal and cardiovascular functions, relieving fatigue, or improving immunity.

In a fifth aspect, the present disclosure provides a microbial agent comprising a kluyveromyces marxianus thallus according to any one of the preceding embodiments, or a composition obtained by a method according to any one of the preceding embodiments.

The inventor tries to use Kluyveromyces marxianus thallus as a main microbial component for preparing biological medicines, and verifies that various different Kluyveromyces marxianus strain thallus can meet the application of the microbial medicines, and the specific realized medicinal effects comprise obviously reducing anaphylactic reaction, effectively regulating intestinal and cardiovascular functions, relieving fatigue and recovering physical stamina and the like.

Based on the above functional application of kluyveromyces marxianus, the present disclosure further provides a composition containing kluyveromyces marxianus for improving compliance of patients taking the composition provided by the present disclosure, the composition contains at least two components with typical recognition smell, and the composition after the components with typical recognition smell are compounded with kluyveromyces marxianus has a new typical recognition smell different from any component in the composition, so that the contradiction emotion of the patient or the eater to the peculiar smell generated by kluyveromyces marxianus is successfully reduced or eliminated.

Still further, the inventors have unexpectedly found that compounding yeast cells (e.g., KM powder) in combination with a nut-flavored, e.g., cashew powder and a spicy-flavored garlic powder can result in a cheese-like flavor composition.

The invention also provides a preparation method of the composition, wherein the main components are solid, and the composition which can be used for preparing the microbial medicine and is provided by the invention can be obtained through grinding and uniformly mixing, and the method is simple, convenient and quick and is suitable for industrial production. Based on this, the present disclosure still further provides a microbial agent comprising kluyveromyces marxianus as a main component, which can be used for preventing allergy, for regulating intestinal and cardiovascular functions, or for relieving fatigue.

Drawings

In order to more clearly illustrate the embodiments of the present disclosure or the prior art, the drawings that are required in the detailed description or the prior art will be briefly described, it will be apparent that the drawings in the following description are some embodiments of the present disclosure, and other drawings may be obtained according to the drawings without inventive effort for a person of ordinary skill in the art.

FIG. 1 is a graph showing the measurement of the water content of the composition obtained in each batch in example 1;

FIG. 2 is a schematic view of the non-caking composition obtained in example 1;

FIG. 3 is a schematic view of the severe caking composition obtained in example 1;

FIG. 4 is a schematic diagram of the slightly agglomerated composition obtained in example 1;

FIG. 5 is the volunteer evaluation result of step S1 of taste evaluation in example 4;

FIG. 6 is the volunteer evaluation result of step S2 of taste evaluation in example 4;

FIG. 7 is the volunteer evaluation result of step S1 of taste evaluation in example 6;

fig. 8 is the volunteer evaluation result of step S2 of taste evaluation in example 6.

Detailed Description

For the purposes of making the objects, technical solutions and advantages of the embodiments of the present disclosure more apparent, the technical solutions of the embodiments of the present disclosure will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present disclosure, and it is apparent that the described embodiments are some embodiments of the present disclosure, but not all embodiments.

The term "typically recognized smell" means that the smell has a commonly recognized, well-defined source to those skilled in the art or to the average consumer, such as various fruit flavors, milk flavors, various flavoring flavors, and the like.

The term "cashew taste" refers to the taste of cashew fruits, cashew (Anacardium occidentale l.) belonging to the genus cashew of the family anaceae, being a angiosperm or small arbor plant, small branch yellow brown, hairless or near hairless; she Gezhi is in the shape of an inverted oval, and the lateral veins are protruded on both sides; the flowers are in a needle-like shape, the flowers are yellow, the sepal is needle-like, and the petals are in a needle-like shape; the fruit support is bright yellow or purple red, and the fruit is kidney-shaped; the flowering period is 12-5 months; the nut is kidney-shaped and is named cashew nut in the fruit period of 4-7 months. Cashew fruits have a typical identified smell, to the extent that they can be easily identified by the average consumer.

The term "walnut taste" refers to the taste of walnut kernels, and the variety of walnuts is varied, including walnuts, iron walnuts, wild walnuts, hemp walnuts, black walnuts, jibao walnuts, heart shaped walnuts, and the like. The "walnut taste" described in the present disclosure does not refer specifically to the taste of a certain walnut kernel because there is a difference in taste between edible nuts of different species of walnut, and it is impossible for a general consumer to accurately distinguish each walnut variety according to taste or smell, but the taste of the walnut as a whole can be determined, and therefore, the present disclosure refers to the taste of such nuts as "walnut taste".

The term "chestnut taste" refers to the taste of chestnut seeds, similar to walnut, and the market of chestnut is numerous, in general, northern chestnut, southern chestnut and japanese chestnut are all popular, and the common chestnut in the market is usually named according to picking areas, such as Qianxi chestnut, qinglong chestnut, chen table chestnut, etc., and the chestnut in different areas has slight differences of waxy feel, soft feel, etc. in terms of taste, while the chestnut in different areas has uniform smell, can be widely known to consumers and clearly distinguished.

The term "pistachio" refers to the taste of pistachio (Pistacia vera l.) fruit, the shape of a ripe pistachio shell split like a smile, is very distinctive in many nuts and even foods, making it highly recognizable in consumers, while also making pistachio taste very recognizable.

The term "macadamia nut taste" refers to the taste of macadamia nut fruits, the original place of the macadamia nut is macadamia nut (macadamia), also called macadamia nut, the aliases are macadamia nut, macadamia nut and the like, and the macadamia nut fruits are widely popularized and planted, so that the macadamia nut is widely known as macadamia nut, and the macadamia nut is characterized by high oil content, various nutrients, and is known as dried fruit queen, and the rich oil content of the macadamia nut enables the macadamia nut to have unique taste and be easily identified by general consumers.

The term "almond taste" refers to the taste of dried mature seeds of the rosaceae plant Prunus armeniaca l.var. The almond is flat, has the length of 1-1.9 cm, the width of 0.8-1.5 cm and the thickness of 0.5-0.8 cm. The surface is yellow brown to dark brown, one end is sharp, the other end is blunt and round, the thickness is thick, and the left and right sides are asymmetric. One side of the tip is provided with a short line-shaped hilum, and a plurality of dark brown veins are upwards arranged at the point of the round end. Thin seed coat, dicotyledon, milky white, oil-rich. No odor, bitter taste, small toxicity, and can be used as medicine for preparing dishes, which is easy to be identified by common consumers.

The term "almond flavor" refers to the flavor of almond, flat and flat, peach-like, non-peach-like, apricot-like, non-apricot-like, very peculiar, so that the name almond is that almond, which can be used as a valuable raw material in the confectionery, pastry, pharmaceutical and cosmetic industries, can be roughly classified into bitter, sweet, soft-shelled sweet almond, and can be easily recognized by general consumers, depending on the sweetness of the kernel flavor.

The term "pine nut taste" refers to the taste of the seed of the Pinaceae plant. Pine nuts, also known as pine cone, pine nut, sea pine nut, are one of the common nuts. Pine nuts are rich in fat, protein, carbohydrate and the like, are important traditional Chinese medicines, are long-term eaten, strengthen body and heart, moisten skin, prolong life and have high dietary therapy value, are instant dried fruits favored by many consumers, are also often used for cooking dishes, and can be easily tasted and identified by general consumers.

The term "Ginkgo taste" refers to the taste of dried mature seeds of Ginkgo biloba l, a plant of the family ginkgaceae, generally the Ginkgo is harvested in autumn when the seeds are ripe, the outer seed coats of the pulp are removed, and the Ginkgo is washed, steamed slightly or boiled slightly, dried and then eaten or used as a medicine. The gingko is slightly elliptical, one end is slightly pointed, the other end is blunt, the length is 1.5-2.5 cm, the width is 1-2 cm, the thickness is about 1cm, the surface of the gingko is yellowish-white or yellowish-brown, smooth and has 2-3 ridge lines; the middle layer is seed coat, is bone and is hard; the inner layer is of seed membranous material, the kernel is wide in oval sphere or ellipse, one end is light brown, the other end is golden yellow, the cross section is yellow, the colloid is like, the inner layer is light yellow or light green, the powder is powdery, and a gap is formed in the middle. No smell, sweet and slightly bitter taste, and can be easily recognized by general consumers.

The term "lotus seed taste" refers to the taste of the mature seed of lotus, a plant of the family Nymphaeaceae. The lotus seeds are slightly elliptic or spheroid, and have a length of 1.2-1.8 cm and a diameter of 0.8-1.4 cm. The surface is light yellow brown to reddish brown, and has fine longitudinal lines and wider veins. One end is nipple-shaped with dark brown center and many cracks, and the periphery is slightly depressed. Hard, thin seed coat and difficult peeling. Dicotyledonous, huang Baise, hypertrophic, with a void in the middle, has green plumula Nelumbinis. No odor, sweet and slightly astringent taste; lotus plumule is bitter in taste, and can be easily tasted and identified by general consumers.

The term "peanut taste" refers to the taste of the seeds of the leguminous plant, the peanut is usually picked up in the end of autumn, the shell is peeled off, and the seeds are sun-dried, commonly known as peanuts. Is one of the most common snack dried fruits, and has a taste which is known to women and children.

The term "chickpea taste" refers to the taste of seeds of the leguminous plant chickpea (Cicer arietinumLinn.) genus. The chickpea seed has complete nutrition and high content. The granule contains 23.0% protein, 63.5% carbohydrate and 5.3% fat. In addition, it is rich in dietary fiber, trace elements and vitamins, and its taste is well known to consumers.

The term "sunflower seed taste" refers to the natural taste of the fruit of sunflower, a plant of the Compositae family. After the sunflower autumn fruits are ripe, cutting the flower disc, sun-drying, beating the fruits, and sun-drying to obtain sunflower seeds, shallow gray or black thin fruits, prolate oval or elliptic, 1 sunflower seed is built in, and the sunflower seeds are light yellow. The sunflower seeds sold in the market can obtain sunflower seed products with various tastes through different manufacturing processes, the sunflower seed taste refers to the original sunflower seed taste, and common consumers can distinguish the original sunflower seeds by tasting before purchasing.

The term "garlic flavor" is the flavor of the allicin released from garlic and sulfides in further metabolites of allicin, which is consistent with the garlic flavor well known in the daily life of the average consumer, and has a very strong recognition.

The term 'green Chinese onion taste', the green Chinese onion is herb of the genus allium of the family liliaceae, contains volatile oil with pungent smell and capsaicin, can remove peculiar smell in greasy and thick-taste dishes such as fishy smell and the like, and generates special aroma. Meanwhile, the allyl sulfide can stimulate gastric secretion, is beneficial to appetite promotion, and can be clearly judged by general consumers.

The term "onion taste" is a biennial herb of the genus allium of the family lycoridae, and the daily consumption of "vegetable onions" is the bulb of onion, the volatile oil in which has a special pungent taste, but which gradually diminishes after heating and also produces a light sweet taste. At the same time, the cell disruption of onion releases alliinase, which reacts with amino acids in onion cells to produce thiopropionaldehyde-S-oxide. The thiopropionaldehyde-S-oxide has volatility, can be released into the air quickly, stimulates free nerve endings on the cornea when contacting eyes, generates burning sensation and stimulates the lacrimal gland to flow out of the lacrimal mouth feel, so that onion flavor can be easily identified for general consumers.

The term cheese or cheese refers to a concentrated dairy product prepared by fermenting animal milk with lactobacillus or adding enzyme, and removing whey, and has sweet taste and milk fragrance, which can be easily identified by common consumers.

In a specific embodiment, the present disclosure provides, in a first aspect, a composition comprising kluyveromyces marxianus, the composition comprising at least two components having a characteristic recognition odor, and the composition after the components having the characteristic recognition odor are compounded with kluyveromyces marxianus having a characteristic recognition odor different from any of the components of the composition.

It is easy to understand that the typical recognition smell after the compounding is a smell which can be generally accepted by a person skilled in the art, general consumers or patients, and does not cause new contradiction emotion, or is a smell generally liked by general consumers, so that the taking compliance is further improved.

It should be noted that, due to the limited biological materials, the present disclosure only examines the feasibility of the 16 kluyveromyces marxianus as described above, which the applicant has deposited by himself, in preparing microbial drugs, and as a result, both can achieve the corresponding physiological functions, and both can successfully achieve flavoring through the composition provided by the present disclosure. Based on this, it will be readily apparent to those skilled in the art that the compositions provided by the present disclosure are capable of achieving the same flavoring effect for other Kluyveromyces marxianus that do not have a particular off-flavor, including but not limited to Kluyveromyces marxianus NBRC 1963, kluyveromyces marxianus DSMZ 7239, kluyveromyces marxianus MTCC 1288, kluyveromyces marxianus IMB3, kluyveromyces marxianus ZIM 1867, kluyveromyces marxianus CBS 6556, kluyveromyces FII 510700, kluyveromyces marxianus ATCC10022 (CBS 7894), kluyveromyces marxianus CDBB-L278, kluyveromyces marxianus NBRC 0219, kluyveromyces marxianus NBRC 0541, kluyveromyces Marxianus NBRC 0617, kluyveromyces NBRC 1777, kluyveromyces marxianus KM1, kluyveromyces BKM-719, kluyveromyces marxianus ATCC10024, kluyveromyces NRRL-Y-2415, kluyveromyces marxianus NRRL-Y-1109, kluyveromyces marxianus Kluyveromyces NRY-610.

In an alternative embodiment, the composition contains 10-20 parts by mass of Kluyveromyces marxianus thallus powder, including but not limited to 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts or 20 parts. The nutty component 74.4-84.7 parts including, but not limited to, 74.4 parts, about 75 parts, about 76 parts, about 77 parts, about 78 parts, about 79 parts, about 80 parts, about 81 parts, about 82 parts, about 83 parts, about 84 parts, or about 84.7 parts. Spicy Xin Liaowei components 0.8-1.2 parts including but not limited to 0.8 parts, 0.9 parts, 1.0 parts, 1.1 parts or 1.2 parts. Wherein the nut-flavored component is preferably cashew dust, the cashew dust comprises a fried cashew dust comprising palm-oil-fried cashew dust and/or coconut-oil-fried cashew dust; the spicy Xin Liaowei component is preferably garlic powder.

The inventors have unexpectedly found that the composition obtained by compounding the fried cashew powder, the garlic powder and the Kluyveromyces marxianus powder turns into a cheese-like taste at the entrance, thereby remarkably improving compliance and having popularization potential.

In an alternative embodiment, the composition further comprises a salt.

The palm oil fried cashew nut powder refers to a method that cashew nuts are put in heated palm oil to be fried to cooked food in a cooking sense, and then processed into powder, wherein the frying temperature is generally 120-150 ℃, including but not limited to 120 ℃, 125 ℃, 130 ℃, 135 ℃, 140 ℃, 145 ℃ or 150 ℃, and the frying time is generally 1-5 min, including but not limited to 1min, 2min, 3min, 4min or 5min.

The coconut oil fried cashew nut powder refers to the method that cashew nuts are placed in heated coconut oil to be fried to cooked food in the cooking sense, and then the cashew nuts are processed into powder, wherein the frying temperature is generally 120-150 ℃, including but not limited to 120 ℃, 125 ℃, 130 ℃, 135 ℃, 140 ℃, 145 ℃ or 150 ℃, and the frying time is generally 1-5 min, including but not limited to 1min, 2min, 3min, 4min or 5min.

The powder essence and the liquid essence added in the present disclosure are cheese-flavored or cheese-flavored essence, and the functions of the essence are as follows: the flavor is increased and improved without affecting the overall odor of the composition.

Strains for use in the present disclosure include:

(1) Kluyveromyces marxianus CJA0001 is preserved in China center for type culture Collection with a preservation number of CCTCC No: m20211601, the preservation date is 2021, 12 and 13, the preservation address is university of Wuhan in Wuhan, china, and the classification name is Kluyveromyces marxianus CJA0001.

(2) Kluyveromyces marxianus CJA0002 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211602, the preservation date is 2021, 12 and 13, the preservation address is Kluyveromyces marxianus CJA0002, classified and named as university of Wuhan in Wuhan, china.

(3) Kluyveromyces marxianus CJA0003 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211603, the preservation date is 2021, 12 and 13, the preservation address is Kluyveromyces marxianus CJA0003.

(4) Kluyveromyces marxianus CJA0004 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211604, the preservation date is 2021, 12 and 13, the preservation address is university of Wuhan in Wuhan, china, and the classification name is Kluyveromyces marxianus CJA0004.

(5) Kluyveromyces marxianus (K. Marxinus) CJA0005, which is preserved in China center for type culture Collection, with the preservation number of CCTCC No: m20211605, the preservation date is 2021, 12 and 13, the preservation address is university of Wuhan in Wuhan, china, and the classification name is Kluyveromyces marxianus CJA.

(6) Kluyveromyces marxianus CJA0006 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211606, the preservation date is 2021, 12 and 13, the preservation address is Kluyveromyces marxianus CJA0006 in class of university of Wuhan in Wuhan, china.

(7) Kluyveromyces marxianus CJA0007 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211607, the preservation date is 2021, 12 and 13, the preservation address is Kluyveromyces marxianus CJA0007 of university of Wuhan in Wuhan, china.

(8) Kluyveromyces marxianus CJA0008 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211608, the preservation date is 2021, 12 and 13, the preservation address is Kluyveromyces marxianus CJA0008 of university of Wuhan in Wuhan, china.

(9) Kluyveromyces marxianus CJA0009 is preserved in China center for type culture Collection, and the preservation number is CCTCC No: m20211609, the preservation date is 2021, 12 and 13, the preservation address is Kluyveromyces marxianus CJA0009 of university of Wuhan in Wuhan, china.

(10) Kluyveromyces marxianus CJA0010 is preserved in China center for type culture Collection (CCTCC No: m20211610, the preservation date is 2021, 12 and 13, the preservation address is university of Wuhan in Wuhan, china, and the classification name is Kluyveromyces marxianus CJA0010.

(11) Kluyveromyces marxianus CJ3113, deposited with China center for type culture Collection with a deposit number of CCTCC No: m20211265, the preservation date is 2021, 10 and 13, the preservation address is university of Wuhan in Wuhan, china, and the classification name is Kluyveromyces marxianus CJ3113.

Some embodiments of the present disclosure are described in detail below with reference to the accompanying drawings. The following embodiments and features of the embodiments may be combined with each other without conflict.

The evaluation method related to the embodiment comprises the following steps:

1. the evaluation method of the dispersibility comprises the steps of passing the composition through a 20-mesh sieve, and counting the influence of the proportion of each component on the caking condition, wherein the condition that the mass of the 20-mesh part of the composition is less than or equal to 50% is serious caking; the mass of the part of the composition with the particle size less than or equal to 20 meshes is between 20 and 50 percent, and the part is medium caking; the part of the composition with the mass less than or equal to 20 meshes is slightly agglomerated when the mass is 5% -20%; the composition has no caking under the condition that the mass of the part less than or equal to 20 meshes is less than 5 percent.

2. The method of evaluating the water content is described in GB/T5009.3-2016 as a direct drying method.

3. The evaluation method of the mouthfeel comprises the steps of requesting volunteers to taste the prepared composition under the double-blind condition, grading oily and sticky mouthfeel, wherein the oily and sticky mouthfeel are respectively provided with 1-10 scores from high to low, the sticky mouthfeel is respectively provided with 1-10 scores from strong to weak, and the average scoring of the volunteers is used as an evaluation result.

4. The taste evaluation method comprises the following two steps:

s1, under double-blind conditions, the volunteers were asked to taste the composition obtained in each batch of experiments, and the results of blind guess taste sources were given.

S2, selecting the first three sources which are guessed most frequently in the step S1 and a composition source summary table consisting of cashew, yeast powder, garlic, walnut, chestnut, pistachio, hawaii nut, almond, pine nut, gingko, lotus seed, peanut, sunflower seed, garlic, green onion and onion, requesting the volunteer to taste each batch of the composition again, and selecting the most probable source, namely the source with the closest taste, from the source summary table.

5. The microorganism detection method comprises the following steps:

the detection method of microorganisms in the fried cashew nuts refers to the detection of the total number of microorganisms in GB 4789.1-2016 food.

Remarks: volunteers described in the above evaluation methods were derived from applicant's internal staff, 49 total, of which men 26 and women 23 were from market sector 8, sales sector 7, administration sector 6 and research and development sector 28, respectively, and none of the volunteers involved the development of the compositions of the present disclosure.

Example 1

In this example, a composition containing different mass fractions of Kluyveromyces marxianus dry powder was prepared using raw cashew, kluyveromyces marxianus dry powder (KM powder) with a preservation number of CCTCC No. M20211265, garlic powder and salt.

The preparation method of the yeast cell dry powder of Kluyveromyces marxianus comprises the following steps: kluyveromyces marxianus with a preservation number of CCTCC No. M20211265 was cultured in a medium (4.5% glucose, 0.2% molasses, 0.2% corn steep liquor dry powder, 0.02% magnesium sulfate, 0.6% ammonium sulfate, 0.6% yeast extract, 0.6% potassium dihydrogen phosphate, 6ppm copper sulfate, 12ppm ferrous sulfate, 15ppm manganese sulfate, 6ppm vitamin B12, 20ppm zinc sulfate, 8ppm methionine, 5ppm alanine, 2ppm cysteine, 5ppm glycine) containing a carbon source, a nitrogen source, and salts; regulating the pH of the culture medium to 4.0-5.0, fermenting at 25-30 ℃ for 40h, heating at 100 ℃ for inactivating for 20min, centrifuging to remove supernatant, obtaining Kluyveromyces marxianus slurry, and spray drying to obtain Kluyveromyces marxianus dry powder.

The garlic powder is non-instant garlic powder sold by Jilin Beijing-extract food Co., ltd and with the number of Q/JSWTM 001S.

The proportions of the components in the composition containing the KM powder in different proportions are shown in Table 1:

table 1 three groups of raw cashew nuts are combined in mass fraction ratio

The preparation method of the composition comprises the following steps: raw cashew nuts and KM powders were weighed according to mass fractions of the compositions in table 1, and then mixed and placed in a knife mill and stirred at a stirring speed of 1400 rpm for a certain period of time to obtain a mixture of KM powders and cashew nut powders, and then garlic powders and salt were added to obtain compositions, and different stirring times were set in each composition according to the following table.

Table 2 mixing time schedule for different ratio compositions of raw cashew nut powder

By controlling the moisture content in the food, the microorganism loses the moisture environment depending on survival, thereby being capable of more conveniently and effectively controlling the growth of the microorganism, preventing the food from caking and deteriorating and prolonging the shelf life of the food. The water content of the composition obtained in each batch was examined by referring to the direct drying method in GB/T5009.3-2016, and as shown in FIG. 1, there was no significant difference between the 9 groups, and the water content was below 5%, which proves that the water content of the composition was not significantly affected by the different KM addition amounts and the different stirring times.

The dispersibility and mouthfeel of each batch of the compositions in table 2 were evaluated with reference to the methods for evaluating dispersibility and mouthfeel of the compositions described above, as follows:

TABLE 3 evaluation results of dispersibility and mouthfeel of compositions of each batch

Remarks: the oil consumption is 10 minutes without oil consumption, 9~7 oil consumption is weak, 6~4 oil consumption is obvious, and 3-1 oil consumption is obvious; the sticky mouthfeel is 10 minutes, the non-sticky mouthfeel is 9~7, the slightly sticky mouthfeel is 6~4, the sticky mouthfeel is more sticky, and the sticky mouthfeel is 3-1.

Example 2

In this example, the raw cashew nuts in example 1 were replaced with cooked cashew nuts prepared by frying raw cashew nuts with 24-degree palm oil at 125 ℃ for 30s, and the rest of the experimental design was the same as that of example 1.

The evaluation results of the obtained composition corresponding to the experimental batch in example 1 were as follows:

table 4 experimental evaluation results of cooked cashew nuts

Note that: the oil consumption is 10 minutes without oil consumption, 9~7 oil consumption is weak, 6~4 oil consumption is obvious, and 3-1 oil consumption is obvious; the sticky mouthfeel is 10 minutes, the non-sticky mouthfeel is 9~7, the slightly sticky mouthfeel is 6~4, the sticky mouthfeel is more sticky, and the sticky mouthfeel is 3-1.

From the experimental results of examples 1 and 2, it can be seen that the content of KM powder and the stirring speed significantly affect the dispersibility of the composition, and the overall oil consumption of the composition prepared by using raw cashew is significantly lower than that of the composition prepared by using cooked cashew, and the overall sticky taste is slightly lower than that of the composition prepared by using cooked cashew.

Example 3

3.1 In this example, on the basis of example 2, the influence of different cooked cashew preparation methods on the stability of the obtained composition was examined, wherein the composition comprises 84.7% of cooked cashew, 10% of KM powder, 1% of garlic powder and 4.6% of salt by mass fraction, the cooked cashew and KM powder were weighed according to mass fraction, then mixed and placed in a knife mill at a stirring speed of 1400 rpm for 60s to obtain a mixture of KM powder and cashew powder, and then garlic powder and salt were added to obtain the composition. The preparation methods of the cooked cashew nuts in different batches are shown in the following table:

table 5 comparative table for different cooked cashew dust preparation methods

3.2 The stability of the composition of different batches obtained in the step 3.1 is examined, the stability detection method comprises the steps of exposing and storing the obtained composition at different temperatures for different times, comparing the smell of the composition with that of the composition obtained by the new preparation, and detecting the microorganism content in the composition after the experiment is finished according to the detection method of the total number of microorganisms in GB 4789.1-2016 food, wherein the specific exposure time and the experiment result are as follows:

TABLE 6 stability test results

Note that: in the scoring of cashew nuts, 10-8 points indicate that the cashew nuts are rich in aroma, 7~4 points indicate that the cashew nuts are weak in aroma, and 3-1 points indicate that the cashew nuts are free of aroma. In the fuel consumption score, 10 parts represent no fuel consumption, 9~7 represents weaker fuel consumption, 6~4 represents more obvious fuel consumption, and 3-1 represents more obvious fuel consumption.

Examples 1 and 2 demonstrate that the oil consumption of the composition prepared from raw cashew nuts is significantly lower than that of cooked cashew nuts, but from the above table, it can be seen that the composition prepared from raw cashew nuts has stability defects, microorganisms attached to raw cashew nuts can be enriched along with the prolongation of the preservation time, the taking compliance of patients is seriously affected, meanwhile, the enriched microorganisms are easy to induce digestive tract diseases of patients, and too high temperature for too long time can cause the overweight of cooked cashew nuts, loss of the aroma of cashew nuts and influence on the flavor of the composition. Thus, when cooked cashew nuts are selected as the raw material component of the composition, the oil temperature is selected to be 120 ℃ and the frying time is 30s, and the temperature can not only remove microorganisms attached to the raw cashew nuts, but also keep the fragrance of the cashew nuts, and the type of the oil used is preferably purified coconut oil. The optimal component proportions in the composition are 74.4% of cooked cashew nuts, 20% of KM powder, 1% of garlic powder and 4.6% of salt by combining the previous experiments. The cooked cashew nuts are prepared by frying purified coconuts, and the oil temperature is 120 ℃ and the frying time is 30s. Weighing cooked cashew nuts and KM powder according to mass fraction, mixing, placing in a knife mill, stirring at 1400 rpm for 60s to obtain a mixture of KM powder and cashew nut powder, and adding garlic powder and salt to obtain the composition.

Example 4

This example the preferred method of preparation of the composition obtained in example 3 was repeated, and the taste of the composition obtained was evaluated using the taste evaluation method described above, and the results and amounts of source guesses given by volunteers in step S1 and step S2 of taste evaluation were shown in fig. 5 and 6, respectively. In step S1 of the double-blind experiment, the blind-guessed taste sources of the volunteers were totally 5, namely cheese, cashew, badam, mushroom and garlic, and the remaining taste sources were all commonly accepted tastes of general consumers or patients except garlic. Among them, cheese (14 persons), cashew (11 persons) and mushroom (9 persons) are considered to be the most taste sources in this order. In step S2, after informing the volunteers of the cheese, cashew, and mushroom and possible raw material taste list, the number of people who guessed the cheese source is still the largest among the taste source guesses that the volunteers again tasted.

Example 5

In order to further improve the mouthfeel and flavor of the composition, cheese flavor is additionally added to the composition obtained in example 4, and the proportions of the components are adjusted as follows:

the powder essence is natural cheese powder essence sold by Guangzhou Yahe biotechnology Co., ltd and having a product number of MZ6600, and the liquid essence is International spice (China) Co., ltd and having a product number of SC1129351 Cheddar cheese essence.

Example 6

The raw cashew nuts used in the preparation method of the preferred composition obtained in example 3 were replaced with chickpeas to obtain a composition comprising chickpeas and KM powder as main components, and the composition was subjected to taste evaluation by the above-mentioned double-blind taste evaluation method, and the results and amounts of source guesses given by volunteers in the S1 and S2 steps of taste evaluation are shown in fig. 7 and 8, respectively. In step S1 of the double blind experiment, the blind guessed taste sources of the volunteers were totally 5, namely cheese, chick pea, almond, mushroom and garlic, and the remaining taste sources were all commonly accepted tastes of general consumers or patients except garlic. Among them, chickpeas (14 persons), cheeses (10 persons) and mushrooms (9 persons) are considered to be the most taste sources in this order. In step S2, after notifying the volunteers of the list of possible raw material tastes of chick peas, cheeses and mushrooms, the number of the chick peas is still the largest among the guessed results of the taste sources that the volunteers taste again, and the number of the yeast powder is zero, which also means that chick peas have the characteristic of masking the yeast flavor, but the taste masking results are more prone to the taste of chick peas themselves.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present disclosure, and not for limiting the same; although the present disclosure has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present disclosure.

Claims

1. The composition containing the Kluyveromyces marxianus thallus is characterized by being a composition formed by compounding a nut taste component, a spicy Xin Liaowei component and the Kluyveromyces marxianus thallus;

the nut flavor component is cashew nut powder, and the spicy Xin Liaowei component is garlic powder.

2. The composition of claim 1, wherein the kluyveromyces marxianus comprises at least one of a preservation number of cctccc No. M20211265, cccc No. M20211601, cccc No. M20211602, cccc No. M20211603, cccc No. M20211604, cccc No. M20211605, cccc No. M20211606, cccc No. M20211607, cccc No. M20211608, cccc No. M20211609, or cccc No. M20211610.

3. The composition of claim 1, wherein the composition comprises, by mass, 10-20 parts of kluyveromyces marxianus thallus powder, 74.4-84.7 parts of cashew powder and 0.8-1.2 parts of garlic powder.

4. The composition of claim 3, wherein the cashew dust comprises a fried cashew dust comprising palm oil fried cashew dust and/or coconut oil fried cashew dust.

5. The composition of claim 4, wherein the composition further comprises a salt.

6. The composition of claim 4, wherein the palm oil is selected from 58 degrees palm oil, 52 degrees palm oil, 44 degrees palm oil, 33 degrees palm oil, or 24 degrees palm oil;

the coconut oil is selected from purified coconut oil or cold pressed coconut oil.

7. The composition of claim 4, wherein the frying temperature is 120-150 ℃ and the frying time is 1-5 min.

8. The composition according to any one of claims 1 to 7, wherein the composition further comprises a powder flavour and/or a liquid flavour;

according to mass fraction, the powder essence is 0.2-0.3 part, and the liquid essence is 0.1-0.15 part;

the aroma of the powder essence and the liquid essence is selected from cheese flavor or cheese flavor.

9. The method for preparing the composition according to any one of claims 1 to 8, wherein the method comprises mixing cashew powder and garlic powder with kluyveromyces marxianus thallus powder, and crushing into powder.

10. Kluyveromyces marxianus thallus, or the composition containing the Kluyveromyces marxianus thallus according to any one of claims 1-8, or the application of the composition obtained by adopting the preparation method according to claim 9 in preparation of microbial medicines.

11. The use according to claim 10, wherein the microbial agent is used for preventing allergies, aiding in the treatment of helicobacter pylori, reducing cholesterol, alleviating diarrhea and enteritis, regulating intestinal and cardiovascular functions, alleviating fatigue, or improving body immunity.

12. A microbial agent comprising kluyveromyces marxianus, or a composition comprising kluyveromyces marxianus according to any one of claims 1 to 8, or a composition comprising kluyveromyces marxianus obtained by the method of claim 9.