阅读说明：本技术 一种基于提高赛门苷含量而提高罗汉果甜苷整体甜度的方法 (Method for improving overall sweetness of mogroside based on improvement of siamenoside content ) 是由 刘晴云 苏志鹏 陈宏坤 于 2019-10-31 设计创作，主要内容包括：本发明公开了一种基于提高赛门苷含量而提高罗汉果甜苷整体甜度的方法,将鲜罗汉果或干罗汉果经提取、树脂吸附、脱色等步骤制得含罗汉果甜苷V含量大于50%的罗汉果甜苷产品,将罗汉果甜苷产品溶于同浓度的发酵培养液中,灭菌处理后,再将原液接种筛选出的特异性清酒酵母菌CN01,自然发酵72h后,可将80%以上的罗汉果苷V转变为赛门苷,离心过滤,收集除去菌体的含高含量赛门苷的发酵液,将此发酵液经离子型色谱树脂分步纯化,收集赛门苷富集段,并经浓缩,干燥制得成品。本发明提高了罗汉果甜苷的附加值,扩展了罗汉果深加工产品品类,产品可作为新型天然甜味剂。(The invention discloses a method for improving the integral sweetness of mogroside based on the improvement of the siamenoside content, which comprises the steps of extracting, resin adsorption, decoloring and the like of fresh or dried momordica grosvenori to prepare a mogroside product containing more than 50% of mogroside V, dissolving the mogroside product in a fermentation culture solution with the same concentration, sterilizing, inoculating a stock solution to screened specific sake yeast CN01, naturally fermenting for 72 hours, converting more than 80% of mogroside V into siamenoside, centrifugally filtering, collecting a fermentation solution containing high-content siamenoside from which thalli is removed, purifying the fermentation solution step by ion type chromatographic resin, collecting a siamenoside enrichment section, concentrating and drying to obtain a finished product. The invention improves the added value of the mogroside, expands the product types of the deep processing of the grosvenor momordica, and the product can be used as a novel natural sweetener.)

1. A method for improving the integral sweetness of mogroside based on the improvement of the siamenoside content is characterized in that: which comprises the following steps:

1) dissolving: dissolving mogroside in a fermentation culture solution with the same concentration, wherein the concentration of the mogroside is 45-50mg/ml, and sterilizing to obtain a stock solution;

2) screening dominant strains:

2-1) separating different strains from the epidermis of the siraitia grosvenorii, the growth soil of the siraitia grosvenorii and the residues of the siraitia grosvenorii, respectively inoculating each strain into a culture solution taking cellulose and starch as substrates, after culturing for 24 hours, determining dominant strains capable of generating exogenous glycosidase by detecting the concentration of glucose in the substrates, and co-screening to obtain five dominant strains with the serial numbers of Sc001, VSc01, CN01, CN01-A and T-CN00, wherein the five dominant strains are saccharomyces cerevisiae strains;

2-2) respectively inoculating saccharomyces cerevisiae strains Sc001, VSc01, CN01, CN01-A and T-CN00 which can generate exogenous glycosidase into a culture solution taking mogroside V as a substrate, fermenting at normal temperature for 168 hours, comparing the sweetness of the fermentation solution with that of the stock solution every 24 hours, and screening out the strain corresponding to the highest sweetness of the fermentation solution as specific sake yeast CN 01;

3) fermentation: rejuvenating specific sake yeast CN01 by using a potato slant culture medium, inoculating the rejuvenated specific sake yeast CN01 to an MS culture medium for propagation, using the rejuvenated specific sake yeast CN01 as a fermentation seed solution after two-stage propagation, and then inoculating the stock solution obtained in the step 1) to the fermentation seed solution, wherein the inoculation amount is 10-12% of the stock solution, the fermentation temperature is controlled to be 30-32 ℃, and the fermentation is carried out for 72-80 h naturally to obtain a fermentation solution;

4) centrifugal filtration: centrifuging and filtering the fermentation liquor, and collecting supernatant;

5) and (3) chromatographic separation: separating the supernatant with transformed ionic chromatographic resin at a flow rate of 2BV/h and for 2h, and collecting the effluent within 45min-1.2 h;

6) concentration and freeze-drying: vacuum concentrating the above effluent, and freeze drying to obtain mogroside with improved sweetness.

2. The method of claim 1, wherein the overall sweetness of mogroside is increased based on increasing the amount of siamenoside, wherein: the mogroside in the step 1) is a spray-dried product of the momordica grosvenori after extraction, resin adsorption and decoloration, and the mogroside V content is more than 50%.

3. The method of claim 1, wherein the overall sweetness of mogroside is increased based on increasing the amount of siamenoside, wherein: and 4) the centrifugation and filtration operations are as follows: centrifuging the fermentation liquor by a butterfly centrifuge with the speed of 6500-7000 r/min, passing through a fine filtration membrane with the diameter of 1um, and collecting the supernatant.

4. The method of claim 1, wherein the overall sweetness of mogroside is increased based on increasing the amount of siamenoside, wherein: the transformation method of the ionic chromatographic resin in the step 5) comprises the following steps: the 001X 7 cation resin was loaded into a chromatographic column and the resin was washed with 2% HCl in a volume of 4BV, then with distilled water to neutrality, then with 2% sodium hydroxide in a volume of 4BV, then with distilled water to neutrality, and finally transformed into LA with 2% LaCl in a volume of 4B^＋And (3) resin.

5. The method of claim 1, wherein the overall sweetness of mogroside is increased based on increasing the amount of siamenoside, wherein: step 6), the operation of concentration and freeze-drying is as follows: vacuum concentrating the effluent to 10 Baume degree, quick freezing to-50 deg.C, and lyophilizing according to the following lyophilization curve: maintaining the temperature at 50 ℃ below zero for 1h, then heating to 30 ℃ below zero for 2h, continuing to heat to 15 ℃ below zero for 2h, continuing to heat to 0 ℃ for 2h, continuing to heat to 15 ℃ for 2h, continuing to heat to 30 ℃ for 4h, continuing to heat to 50 ℃ for 8h, then cooling to 30 ℃ for 4h, and finally cooling to 25 ℃ for 2 h.

Technical Field

The invention relates to the field of momordica grosvenori processing, and mainly relates to a method for improving the integral sweetness of momordica grosvenori sweet glycosides based on the improvement of the content of siamenoside.

Background

The mogroside is also called mogroside, is extracted from the special economic plant of Guangxi-fructus momordicae, the sweetness of which is 300 times of that of cane sugar, the heat is zero, and the mogroside has the effects of clearing heat, moistening lung, relieving cough, moistening intestines and relaxing bowels, and has the prevention and treatment effects on obesity, constipation, diabetes and the like. However, the actual highest sweetness of the grosvenor momordica is not the mogroside V but the siamenoside, the sweetness of which is about 1.5 times of the mogroside V, the chemical structure of which is just one beta-D-glucose less than the mogroside V, but the content of the siamenoside in the grosvenor momordica is extremely low. If the content of the siamenoside can be increased, the sweetness of the mogroside can be improved as a whole.

Disclosure of Invention

The invention aims to provide a method for improving the integral sweetness of mogroside based on the improvement of the siamenoside content by screening a dominant strain to hydrolyze a beta-D-grape base.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for improving the overall sweetness of mogroside based on the improvement of the siamenoside content comprises the following steps:

1) dissolving: dissolving mogroside in a fermentation culture solution with the same concentration, wherein the concentration of the mogroside is 45-50mg/ml, and sterilizing to obtain a stock solution;

2) screening dominant strains:

2-1) separating different strains from the epidermis of the siraitia grosvenorii, the growth soil of the siraitia grosvenorii and the residues of the siraitia grosvenorii, respectively inoculating each strain into a culture solution taking cellulose and starch as substrates, after culturing for 24 hours, determining dominant strains capable of generating exogenous glycosidase by detecting the concentration of glucose in the substrates, and co-screening to obtain five dominant strains with the serial numbers of Sc001, VSc01, CN01, CN01-A and T-CN00, wherein the five dominant strains are saccharomyces cerevisiae strains;

2-2) respectively inoculating saccharomyces cerevisiae strains Sc001, VSc01, CN01, CN01-A and T-CN00 which can generate exogenous glycosidase into a culture solution taking Morgoroside V as a substrate, fermenting at normal temperature for 168 hours, comparing the sweetness of the fermentation solution with that of the stock solution every 24 hours, and screening out the strain corresponding to the highest sweetness of the fermentation solution as specific sake yeast CN 01;

3) fermentation: rejuvenating specific sake yeast CN01 by using a potato slant culture medium, inoculating the rejuvenated specific sake yeast CN01 to an MS culture medium for propagation, using the rejuvenated specific sake yeast CN01 as a fermentation seed solution after two-stage propagation, and then inoculating the stock solution obtained in the step 1) to the fermentation seed solution, wherein the inoculation amount is 10-12% of the stock solution, the fermentation temperature is controlled to be 30-32 ℃, and the fermentation is carried out for 72-80 h naturally to obtain a fermentation solution;

4) centrifugal filtration: centrifuging and filtering the fermentation liquor, and collecting supernatant;

5) and (3) chromatographic separation: separating the supernatant with transformed ionic chromatographic resin at a flow rate of 2BV/h and for 2h, and collecting the effluent within 45min-1.2 h;

6) concentration and freeze-drying: vacuum concentrating the above effluent, and freeze drying to obtain mogroside with improved sweetness.

The mogroside in the step 1) is a spray-dried product of the momordica grosvenori after extraction, resin adsorption and decoloration, and the Morgoroside V content of the mogroside is more than 50%.

And 4) the centrifugation and filtration operations are as follows: centrifuging the fermentation liquor by a butterfly centrifuge with the speed of 6500-7000 r/min, passing through a fine filtration membrane with the diameter of 1um, and collecting the supernatant.

The transformation method of the ionic chromatographic resin in the step 5) comprises the following steps: the 001X 7 cation resin was loaded into a chromatographic column and the resin was washed with 2% HCl in a volume of 4BV, then with distilled water to neutrality, then with 2% sodium hydroxide in a volume of 4BV, then with distilled water to neutrality, and finally transformed into LA with 2% LaCl in a volume of 4B^＋And (3) resin.

Step 6), the operation of concentration and freeze-drying is as follows: vacuum concentrating the effluent to 10 Baume degree, quick freezing to-50 deg.C, and lyophilizing according to the following lyophilization curve: maintaining the temperature at 50 ℃ below zero for 1h, then heating to 30 ℃ below zero for 2h, continuing to heat to 15 ℃ below zero for 2h, continuing to heat to 0 ℃ for 2h, continuing to heat to 15 ℃ for 2h, continuing to heat to 30 ℃ for 4h, continuing to heat to 50 ℃ for 8h, then cooling to 30 ℃ for 4h, and finally cooling to 25 ℃ for 2 h.

The invention adopts the technical scheme that fresh momordica grosvenori or dry momordica grosvenori is subjected to steps of extraction, resin adsorption, decoloration and the like to prepare a momordica grosvenori sweet glycoside product containing momordica grosvenori sweet glycoside V more than 50%, the momordica grosvenori sweet glycoside is dissolved in fermentation culture solution with the same concentration, after sterilization treatment, the stock solution is inoculated with the screened specific sake yeast CN01, after natural fermentation is carried out for 72 hours, more than 80% of momordica grosvenori glycoside V can be converted into siamenoside, centrifugal filtration is carried out, fermentation liquor containing high-content siamenoside is collected, the fermentation liquor is subjected to step purification by ionic chromatographic resin, a siamenoside enrichment section is collected, and the finished product is prepared by concentration and drying. Momordica grosvenori produces many specific enzymes in the maturation process, and these enzymes are necessary for producing momordica grosvenori sweet glycosides. According to the invention, through fermentation, the production conditions of sweet substances are simulated, low-sweet substances are directionally converted into high-sweet substances, and the additional value of the deeply processed product of the momordica grosvenori is improved.

According to the invention, the mogroside is converted into high-sweetness siamenoside through specific fermentation, the prepared siamenoside has long sweetness, has no aftertaste, is nearly 1.5 times higher in sweetness compared with the same-concentration mogroside, and can be applied to various sugar substitute products and weight-losing health-care foods. The invention improves the added value of the mogroside, expands the product types of the deep processing of the grosvenor momordica, and the product can be used as a novel natural sweetener.

Detailed Description

A method for improving the overall sweetness of mogroside based on the improvement of the siamenoside content comprises the following steps:

1) dissolving: dissolving mogroside with Morgoroside V content of more than 50% obtained by extracting fructus Siraitiae Grosvenorii, adsorbing with resin, decolorizing, spray drying in fermentation culture solution with the same concentration, wherein the mogroside concentration is 45-50mg/ml, and sterilizing to obtain stock solution;

2-1) separating different strains from the epidermis of the siraitia grosvenorii, the growth soil of the siraitia grosvenorii and the residues of the siraitia grosvenorii, respectively inoculating each strain into a culture solution taking cellulose and starch as substrates, after culturing for 24 hours, determining dominant strains capable of generating exogenous glycosidase by detecting the concentration of glucose in the substrates, and co-screening to obtain five dominant strains with the serial numbers of Sc001, VSc01, CN01, CN01-A and T-CN00, wherein the five dominant strains are saccharomyces cerevisiae strains;

2-2) respectively inoculating saccharomyces cerevisiae strains Sc001, VSc01, CN01, CN01-A and T-CN00 which can generate exogenous glycosidase into a culture solution taking Morgoroside V as a substrate, fermenting at normal temperature for 168 hours, comparing the sweetness of the fermentation solution with that of the stock solution every 24 hours, and screening out the strain corresponding to the highest sweetness of the fermentation solution as specific sake yeast CN 01;

3) fermentation: rejuvenating specific sake yeast CN01 by using a potato slant culture medium, inoculating the rejuvenated specific sake yeast CN01 to an MS culture medium for propagation, using the rejuvenated specific sake yeast CN01 as a fermentation seed solution after two-stage propagation, and then inoculating the stock solution obtained in the step 1) to the fermentation seed solution, wherein the inoculation amount is 10-12% of the stock solution, the fermentation temperature is controlled to be 30-32 ℃, and the fermentation is carried out for 72-80 h naturally to obtain a fermentation solution;

4) centrifugal filtration: centrifuging the fermentation liquor by a butterfly centrifuge with the speed of 6500-7000 r/min, passing through a fine filtration membrane with the diameter of 1um, and collecting supernatant;

5) and (3) chromatographic separation: separating the supernatant with transformed ionic chromatographic resin at a flow rate of 2BV/h and for 2h, and collecting the effluent within 45min-1.2 h;

6) concentration and freeze-drying: vacuum concentrating the above eluate under reduced pressure to 10 Baume degree, quick freezing to-50 deg.C, and lyophilizing according to the following lyophilization curve: maintaining the temperature at 50 ℃ below zero for 1h, then heating to 30 ℃ below zero for 2h, continuously heating to 15 ℃ below zero for 2h, continuously heating to 0 ℃ for 2h, continuously heating to 15 ℃ for 2h, continuously heating to 30 ℃ for 4h, continuously heating to 50 ℃ for 8h, then cooling to 30 ℃ for 4h, and finally cooling to 25 ℃ for 2h to obtain the mogroside with improved sweetness.

The strains Sc001, VSc01, CN01, CN01-A and T-CN00 can be obtained from Min food (Zhangzhou) limited company.

The transformation method of the ionic chromatographic resin comprises the following steps: the 001X 7 cation resin was loaded into a chromatographic column and the resin was washed with 2% HCl in a volume of 4BV, then with distilled water to neutrality, then with 2% sodium hydroxide in a volume of 4BV, then with distilled water to neutrality, and finally transformed into LA with 2% LaCl in a volume of 4B^＋And (3) resin.