阅读说明：本技术 侧耳木霉菌株zj-03在工业大麻精深加工中的应用 (Application of trichoderma pleuroticola ZJ-03 in deep processing of industrial hemp ) 是由 钱朋智 张梅娟 于 2020-05-19 设计创作，主要内容包括：侧耳木霉菌株ZJ-03在工业大麻精深加工中的应用,涉及食品加工技术领域。本发明的目的是要解决如何诱导侧耳木霉产高酶活的木聚糖酶,以及如何制备出高纯度且主要成分为木二糖和木三糖的健康糖浆产品的问题。方法：将汉麻屑粉和汉麻籽粕粉加入到营养液中,搅拌,接种侧耳木霉ZJ-03孢子悬液；接种后置于发酵室进行培养,得到侧耳木霉固体发酵料,将其静止浸提,得到浸提固液混合物,将其进行粗过滤,再离心,取上清液,得到粗糖液；将粗糖液依次进行第一次蒸发浓缩、第一次离子交换、第二次蒸发浓缩、第二次离子交换和第三次蒸发浓缩,得到功能糖浆产品。本发明可获得侧耳木霉菌株ZJ-03在工业大麻精深加工中的应用。(Application of trichoderma pleuroticola ZJ-03 in deep processing of industrial hemp relates to the technical field of food processing. The invention aims to solve the problems of how to induce trichoderma pleuroticola to produce xylanase with high enzyme activity and how to prepare a health syrup product with high purity and main components of xylobiose and xylotriose. The method comprises the following steps: adding hemp scrap powder and hemp seed meal powder into the nutrient solution, stirring, and inoculating trichoderma aureoviride ZJ-03 spore suspension; inoculating, culturing in a fermentation chamber to obtain a trichoderma aureobasicola solid fermentation material, standing and leaching to obtain a leaching solid-liquid mixture, performing coarse filtration on the solid-liquid mixture, centrifuging again, and taking supernatant to obtain a coarse sugar liquid; and sequentially carrying out first evaporation concentration, first ion exchange, second evaporation concentration, second ion exchange and third evaporation concentration on the crude sugar solution to obtain a functional syrup product. The invention can obtain the application of the trichoderma aureoviride ZJ-03 in the deep processing of industrial hemp.)

1. The application of trichoderma pleuroticola ZJ-03 in the deep processing of industrial hemp is characterized by comprising the following steps:

inoculating trichoderma aureoviride ZJ-03 spores into a PDA slant culture medium under an aseptic condition, and then placing the culture medium into a mould incubator for culture to obtain activated trichoderma aureoviride ZJ-03 strain spores; taking down the activated spore of the Trichoderma aureoviride ZJ-03 strain, scattering and shaking up to obtainTo a concentration of 1.5 × 10⁷2.0 to 2.0 × 10⁷each/mL of Trichoderma aureoviride ZJ-03 spore suspension;

secondly, adding hemp seed meal powder and hemp crumb powder into the nutrient solution, fully stirring and sterilizing, and then inoculating the trichoderma aureobasicola ZJ-03 spore suspension obtained in the first step under an aseptic condition, wherein the inoculation amount is 1.5-2%; after inoculation, placing the inoculated product in a sterilized fermentation chamber, firstly culturing for 68-72 h at 28-30 ℃, then culturing for 20-24 h at 33-37 ℃, and then continuously culturing for 22-26 h at 37-40 ℃ to obtain a trichoderma aureobasicola solid fermentation material; leaching the trichoderma aureobasicola solid fermentation material to obtain a leached solid-liquid mixture; carrying out coarse filtration and centrifugation on the solid-liquid mixture obtained by leaching, and taking supernatant to obtain a crude sugar solution;

and thirdly, sequentially carrying out first evaporation concentration, first ion exchange, second evaporation concentration, second ion exchange and third evaporation concentration on the crude sugar liquid obtained in the step two to obtain a functional syrup product.

2. The application of trichoderma pleuroticum strain ZJ-03 in the deep processing of industrial cannabis sativa according to claim 1, wherein the hemp sawdust powder in the second step is prepared by the following steps: crushing hemp scraps, sieving with a 40-mesh sieve, and taking undersize products to obtain hemp scrap powder; the hemp seed meal powder is prepared by the following steps: crushing hemp seed meal, sieving with a 40-mesh sieve, and taking undersize to obtain hemp seed meal powder; the nutrient solution is prepared by the following steps: adding the glucose mother liquor and the threonine mother liquor into distilled water, and uniformly mixing to obtain the nutrient solution, wherein the ratio of the mass of the glucose mother liquor to the mass of the threonine mother liquor to the volume of the distilled water is (0.5 g-1 g): (0.5 g-1 g): 100 mL.

3. The use of Trichoderma aureoviride strain ZJ-03 in the intensive processing of industrial cannabis according to claim 1, wherein the cultivation in step one is as follows: culturing at 28-30 ℃ until the spores of trichoderma pleuroticola ZJ-03 are mature; and (2) sterilization in the second step: stirring thoroughly, placing into container, and sterilizing at 121 deg.C for 2 hr; leaching in the second step: adding the trichoderma pleuroticola solid fermentation material into pure water, and statically leaching for 6-8 h at the temperature of 35-37 ℃; centrifuging in the step two: centrifuging the solid-liquid mixture for 10-15 min under 3500-4000 r/min.

4. The application of trichoderma aureobasilicum strain ZJ-03 in the fine and further processing of industrial hemp according to claim 1 or 3, characterized in that the mass ratio of hemp scrap powder to hemp seed meal powder in the second step is (19-49): 1, and the ratio of the total mass of hemp scrap powder and hemp seed meal powder to the volume of the nutrient solution is 1 (1.5-1.8); the ratio of the mass of the trichoderma pleuroticola solid fermentation material to the volume of pure water is 1 g: (8 mL-10 mL).

5. The use of Trichoderma aureoviride strain ZJ-03 in the intensive processing of industrial cannabis according to claim 1, wherein the first evaporative concentration in step three is carried out by the following steps: and (3) adding the crude sugar solution obtained in the step two into a rotary evaporator, and evaporating and concentrating at the temperature of 75-85 ℃ and at the rotating speed of 25-35 r/min until the concentration of soluble solids in the crude sugar solution is 8-12 g/100mL to obtain a first concentrated solution.

6. The use of Trichoderma aureoviride strain ZJ-03 in the intensive processing of industrial cannabis according to claim 1, wherein the first ion exchange in step three is carried out by the following steps: and (3) passing the first concentrated solution through a first anion exchange resin column, a first cation exchange resin column and a second anion exchange resin column in sequence to obtain a first ion exchange solution.

7. The use of Trichoderma aureoviride strain ZJ-03 in the intensive processing of industrial cannabis according to claim 1, wherein the second evaporative concentration in step three is carried out by the following steps: and adding the first ion exchange liquid into a rotary evaporator, and evaporating and concentrating at the temperature of 75-85 ℃ and at the rotating speed of 25-30 r/min until the concentration of soluble solids in the first ion exchange liquid is 10-20 g/100mL to obtain a second concentrated liquid.

8. The use of Trichoderma aureoviride strain ZJ-03 in the intensive processing of industrial cannabis according to claim 1, wherein the second ion exchange in step three is carried out by the following steps: and (3) passing the second concentrated solution through a second cation exchange resin column, a third anion exchange resin column and a third cation exchange resin column in sequence to obtain a second ion exchange solution.

9. The use of Trichoderma aureoviride strain ZJ-03 in the intensive processing of industrial cannabis according to claim 1, wherein the third evaporative concentration in step three is carried out by the following steps: and adding the second ion exchange liquid into a rotary evaporator, and evaporating and concentrating at the temperature of 75-85 ℃ and at the rotating speed of 10-20 r/min until the concentration of soluble solids in the second ion exchange liquid is 65-75 g/100mL, thereby obtaining the functional syrup product.

10. The use of Trichoderma aureoviride strain ZJ-03 in the fine and further processing of industrial hemp according to claim 6 or 8, wherein the type of the resin in the first anion exchange resin column is D301, the type of the resin in the first cation exchange resin column is 001, the type of the resin in the second anion exchange resin column is D201, the type of the resin in the second cation exchange resin column is 001 x 7, the type of the resin in the third anion exchange resin column is D301, and the type of the resin in the third cation exchange resin column is 001 x 7.

Technical Field

The invention relates to the technical field of food processing, in particular to application of trichoderma aureobasilicum strain ZJ-03 in fine and further processing of industrial hemp.

Background

Industrial hemp (hemp), a plant of the genus Cannabis of the family Moraceae, is a hemp variety that has been artificially bred and approved (identified or registered) by crop varieties, has a tetrahydrocannabinol content of less than 0.3% (dry matter percentage) at the top leaves and flower ears of the flowering season of the plant population, and has no drug extraction or direct drug utilization value.

The continuous development of the hemp industry becomes a difficult problem to be solved urgently, modern food mechanical equipment integrates deep processing of the hemp into the food industry, a wider development space is opened up for the hemp, and the value of the hemp is improved while the industrial chain is deepened. In the international market, the types of hemp food products reach more than 3000. The hemp food has rich varieties, wide industry prospect in the fields of catering, health-care food and the like, and great market potential.

The hemp functional components are effectively utilized to develop hemp functional food, health food or medicine, which not only plays an important role in the development of hemp industry, but also is beneficial to the health of people. At present, research and development on hemp functional products have become one of hot industries. But functional products with high added value are still to be researched and developed. China hemp industry needs to gradually improve comprehensive utilization rate, realize international layout and enrich market application, which is the key for improving basic capability and industrial chain level of China hemp industry. The key to realize new breakthrough in the fields of food, medicine, new materials and the like is to further enrich high value-added products taking the intensive extraction of the hemp as raw materials. Wherein, the trichoderma pleuroticola fermentation is applied to the fine and further processing of industrial hemp to prepare syrup, which is not reported yet; in addition, how to produce xylanase with high enzyme activity and how to prepare high-purity health syrup products become technical problems in the food processing industry.

Disclosure of Invention

The invention aims to solve the problems of inducing trichoderma aureoviride to produce xylanase with high enzyme activity and preparing a health syrup product with high purity and main components of xylobiose and xylotriose, and provides application of trichoderma aureoviride ZJ-03 in deep processing of industrial hemp.

The application of the trichoderma pleuroticola ZJ-03 in the fine and further processing of industrial hemp is completed according to the following steps:

firstly, inoculating Trichoderma aureoviride ZJ-03 spores into a PDA slant culture medium under the aseptic condition, then placing the culture medium into a mould incubator for culture to obtain activated Trichoderma aureoviride ZJ-03 strain spores, taking down the activated Trichoderma aureoviride ZJ-03 strain spores, scattering and shaking up to obtain the Trichoderma aureoviride ZJ-03 strain spores with the concentration of 1.5 × 10⁷2.0 to 2.0 × 10⁷each/mL of Trichoderma aureoviride ZJ-03 spore suspension;

secondly, adding hemp seed meal powder and hemp crumb powder into the nutrient solution, fully stirring and sterilizing, and then inoculating the trichoderma aureobasicola ZJ-03 spore suspension obtained in the first step under an aseptic condition, wherein the inoculation amount is 1.5-2%; after inoculation, placing the inoculated product in a sterilized fermentation chamber, firstly culturing for 68-72 h at 28-30 ℃, then culturing for 20-24 h at 33-37 ℃, and then continuously culturing for 22-26 h at 37-40 ℃ to obtain a trichoderma aureobasicola solid fermentation material; leaching the trichoderma aureobasicola solid fermentation material to obtain a leached solid-liquid mixture; carrying out coarse filtration and centrifugation on the solid-liquid mixture obtained by leaching, and taking supernatant to obtain a crude sugar solution;

and thirdly, sequentially carrying out first evaporation concentration, first ion exchange, second evaporation concentration, second ion exchange and third evaporation concentration on the crude sugar liquid obtained in the step two to obtain a functional syrup product.

The invention has the beneficial effects that:

1. the invention realizes that the trichoderma aureoviride ZJ-03 strain takes the hemp chips and the hemp seed meal as main growth substrates for solid state fermentation, and induces the produced xylanase to degrade the hemp chips and the hemp seed meal to be synchronously carried out in the growth period, thereby saving the time and the cost for extracting and refining the xylanase, simplifying the preparation process of the functional syrup product, having feasibility, simultaneously taking the hemp chips and the hemp seed meal as the base material for microbial growth and the acting substrate of the xylanase, fully utilizing the hemp chips and the hemp seed meal, increasing the additional values of the hemp chips and the hemp seed meal, and exploring a new way for the comprehensive utilization of the hemp and the preparation of the functional syrup product.

2. According to the industrial hemp (hemp) functional syrup product, the hemp sawdust powder and the hemp seed meal powder are used as main solid fermentation growth substrates, so that the enzyme activity of xylanase produced by trichoderma pleuroticola is induced to be improved by more than 20% compared with the existing enzyme activity (namely, more functional syrup products with the same purity can be produced by the same amount of xylanase); the impurities such as pigment, protein, monosaccharide and polysaccharide in the functional syrup product are removed by adopting separation and purification technical means such as leaching, coarse filtration and centrifugation, and subsequent multiple evaporation concentration and ion exchange of a coarse sugar solution, so that the purity of the final functional syrup product is as high as 72.80%, and the main components are xylobiose (the purity is as high as 46.00%) and xylotriose (the purity is as high as 26.80%), the main components in the functional syrup product are xylobiose and xylotriose, the products decomposed by digestive enzymes after the xylobiose and the xylotriose are absorbed by a human body are xylose, the xylose is difficult to be decomposed by the digestive enzymes of the human body, saccharides such as glucose and the like which can increase the blood sugar and the blood fat of the human body can not be generated, and the xylose only contains little heat; therefore, the functional syrup product is low-calorie health syrup, and has multiple effects of controlling blood sugar, preventing obesity, improving diabetes symptoms and the like when being added into food.

3. The invention has simple technical route, low cost and good development prospect and market prospect.

The invention can obtain the application of the trichoderma aureoviride ZJ-03 in the deep processing of industrial hemp.

Drawings

FIG. 1 is a flow chart of the process of producing functional syrup products from solid state fermentation of trichoderma pleurorum and separating and purifying functional syrup products according to the embodiment;

FIG. 2 is an activated spore of Trichoderma aureoviride ZJ-03 strain;

FIG. 3 is a schematic diagram of a Trichoderma reesei solid fermentation material;

FIG. 4 is a high performance liquid chromatogram of a functional syrup product.

Detailed Description