CN102334702B - Production method for laver protein and polysaccharide nutrient powder - Google Patents
Production method for laver protein and polysaccharide nutrient powder Download PDFInfo
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- CN102334702B CN102334702B CN201110195046XA CN201110195046A CN102334702B CN 102334702 B CN102334702 B CN 102334702B CN 201110195046X A CN201110195046X A CN 201110195046XA CN 201110195046 A CN201110195046 A CN 201110195046A CN 102334702 B CN102334702 B CN 102334702B
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Abstract
The invention discloses a production method for laver protein and polysaccharide nutrient powder, and relates to laver. The method comprises the following steps of: adding microbial bacterial solution into soaked laver; fermenting with a shaking table to obtain laver fermentation liquor; adding protease into the laver fermentation liquor with adjusted pH for enzymolysis; removing insoluble substances through heating sterilization, enzyme deactivation, centrifugal separation and filtering to obtain laver fermentation enzymolysis solution; and drying the laver fermentation enzymolysis solution to obtain the laver protein and polysaccharide nutrient powder. By combining microbial fermentation and proteolysis, extraction yield of soluble nutritive materials is high, and more than 75g of soluble laver nutrient powder can be obtained from 100g of laver; a new approach is developed for deep processing of laver and product development; the nutrient powder can be used as a natural food additive to be applied to food industry and can also be used as an intermediate raw material to research and develop various novel functional foods; and by using the laver protein and polysaccharide nutrient powder, the laver protein and laver polysaccharide which are further separated and purified can also be applied to industries of biomedicine, cosmetics and the like.
Description
Technical field
The present invention relates to a main laver, especially relate to the production method of a main laver proteoglycan nutriment powder.
Background technology
China's algae resource is very abundant, and wherein laver culture output has reached 110,000 tons.Yet because the laver process technology lags behind, the intensive processing product is few, rests on for a long time on dried product and the roasting laver aspect, and the economic benefit that high yield and high-quality laver produce is not also further deepened.On the other hand, protein content contains abundant essential amino acid up to 25%～50% in the laver, and fat content is low, is rich in the multiple nutritional components such as vitamin, mineral matter, is the desirable feedstock of health food.Along with the quickening of life modernization, the healthy nutritive value of laver also constantly comes into one's own.Yet because the laver cell wall, human body can't fully be digested and assimilated its nutritional labeling, and the nutritive value of laver is not utilized effectively.
For the health nutrient composition that exists in the laver, at present, lot of domestic and international research institution all is certain active material that concentrates in the simple separation laver, but extracting method tends to its active material is damaged or wastes improperly, has affected giving full play to that laver nutritive is worth.At present, the extraction of laver albumen is mainly adopted the wall-breaking methods such as multigelation method, swelling method and impulse ultrasound method, recovery rate is mostly 15%～45%.Cold water extraction method, hot water extraction method are generally adopted in the extraction of laver amylose, also have in addition the methods such as microwave and ultrasound assisted extraction, and recovery rate is generally 15%～35%.Recovery rate is all lower, and the result has also caused achievement in research to apply in practice.
Jiangsu University provides the method for a kind of purple laver protein and polyose and ultrasonic assisted extraction thereof in Chinese patent CN1687122.When preparing purple laver protein/polyose take dried laver as raw material, adopt following technological process: dried laver → pulverize → remove fat → ultrasonic auxiliary water extraction → laver water extract → filtration → concentrated → spray-drying → purple laver protein/polysaccharide powder.The product that obtains contains protein 40%～50%, polysaccharide compound 22%～30%.
Liu Yi discloses the extracting method of a kind of laver amylose and laver albumen in Chinese patent CN102002111A, the method is with after the laver oven dry, pulverizing, and the auxiliary sig water of ultrasonic wave extracts albumen and polysaccharide; The isoelectric point method protein precipitation, ultrafiltration purification, the spray-dried laver protein product that obtains; The residue that will extract again behind the albumen cleans to neutral, and dynamic super-voltage extracts polysaccharide, activated carbon decolorizing, and ultrafiltration purification obtains the laver amylose product after the spray-drying.
The general using chemical reagent is processed in conjunction with the hot water extraction and is extracted the laver proteoglycan, its solid content recovery rate only have 19%～23% (Wu Kegang. the extraction of laver juice and take off raw meat and protect look research. food industry science and technology, 2006,5).And at present for the highest ultrasonic disruption extraction method of laver extract content, the recovery rate of laver amylose and laver albumen also only have 15%～45% (Xiao Haifang. pulse ultrasonic wave extracts the technical study of albumen and polysaccharide in the laver simultaneously. Food Science, 2007,6).
Summary of the invention
The object of the present invention is to provide the production method of a main laver proteoglycan nutriment powder.
Technical scheme of the present invention is to utilize first bacillus subtilis or bafillus natto that water purification is soaked the laver that disperses to carry out shaker fermentation, then to the further proteolysis of laver zymotic fluid, at last the supernatant of centrifugal acquisition utilized freeze drying or spray-drying, be prepared into and have good water-soluble laver proteoglycan nutriment powder.
The present invention includes following steps:
1) preparation laver zymotic fluid: add microbial inoculum in the laver that immersion is opened, shaker fermentation gets the laver zymotic fluid;
In step 1) in, the concrete grammar of described preparation laver zymotic fluid is: first microorganism fungus kind is utilized the LB solid medium at 37 ℃ of lower 8～12h of cultivation, picking is cultivated 16～24h with the LB fluid nutrient medium after going out single bacterium colony under 37 ℃, microbial inoculum as fermentation usefulness, the recycling microbial inoculum carries out shaker fermentation to laver, is prepared into the laver zymotic fluid; Described microorganism fungus kind can be a kind of in bacillus subtilis or the bafillus natto; The temperature of described shaker fermentation can be 30 ℃, and the time of described shaker fermentation can be 12～36h, and the rotating speed of described shaker fermentation can be 120～200rpm; The addition of described microbial inoculum can be 1%～4% of laver aqueous solution volume.
2) enzymolysis of laver zymotic fluid: in adjusting the laver zymotic fluid of pH, add protease hydrolyzed, remove insoluble substance by heat sterilization, the enzyme that goes out, centrifugation, filtration, obtain laver fermentation enzymolysis liquid;
In step 2) in, described pH can be 5.5～6.5; The addition of described protease can be 0.05%～0.2% of laver zymotic fluid by volume, and described protease can adopt papain, and the temperature of described enzymolysis can be 55 ℃, and the time of described enzymolysis can be 2～4h.
3) preparation of laver proteoglycan nutriment powder: with the laver enzymolysis liquid drying of fermenting, namely get laver proteoglycan nutriment powder.
In step 3) in, described drying can adopt freeze drying or spray-drying etc.
Compare with the extracting method of existing laver proteoglycan, the present invention has following outstanding advantages:
1) the general using chemical reagent is processed in conjunction with the hot water extraction and is extracted the laver proteoglycan, its solid content recovery rate only has 19%～23%, and at present for the highest ultrasonic disruption extraction method of laver extract content, the recovery rate of laver amylose and laver albumen also only has 15%～45%.The present invention is owing to utilize microbial fermentation and proteolysis technology destruction laver cell wall, making in the laver nutriment such as albumen, polysaccharide dissociate out from laver cell becomes water-soluble substances, extract yield is high, and technology both can be fit to large-scale industrialization production, can reduce production costs again.
2) bafillus natto and the bacillus subtilis of the present invention's employing all are the safe microorganisms of no pathogenicity, thalline self can cellulase synthesis and the enzyme such as protease, utilize it that laver is fermented, can reduce production costs on the one hand, go on the other hand large-scale industrialization production.
3) laver proteoglycan nutriment powder of the present invention extracts the yield height, and wherein the water-soluble solid content recovery rate of laver is greater than 75%, and the laver protein extracting ratio is greater than 83%.
4) extracting method of the present invention can not destroyed the good protein of laver.
5) in the laver proteoglycan nutriment powder of the present invention's preparation, after tested, protein content reaches 29%～31%, and polyoses content reaches 24%～40%.
6) the laver proteoglycan nutriment powder of the present invention preparation, protein solubility be near 100%, and affected not quite by pH, as the food additives applied range.
7) the present invention also can be used as the pre-treating method of developing Medicines and Health Product and functional food.
8) the invention provides a kind of method of effective destruction laver cell wall, utilize microorganism first the laver cell wall to be destroyed, again with the combination between protease destruction laver amylose and the laver albumen, make that the various nutriments to the human body beneficial can both become water-soluble substances in the laver, reach the effect of high efficiency extraction, extract not only can be used as natural additive for foodstuff, can also research and develop various novel functional foods as intermediate raw material, thereby promote the development of laver deep processing research, drive the sound development of laver culture industry.
The specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment.
Embodiment 1
1, the preparation of bacillus subtilis bacterium liquid: Bacillus subtilis strain is provided by Microbiological Lab of bioengineering institute of Collects The American University, utilize the LB solid medium at 37 ℃ of lower 8h of cultivation, recycling LB fluid nutrient medium was at 37 ℃ of lower 16h of cultivation, for following laver fermentation after picking went out single bacterium colony.
2, the preparation of laver zymotic fluid: get the 8g laver and put into 200ml distilled water, add simultaneously the bacillus subtilis bacterium liquid 4ml of preparation in the step (1), at 30 ℃ of shaker fermentation 24h, be prepared into the laver zymotic fluid.
3, the enzymolysis of laver zymotic fluid: utilize acetic acid that the laver zymotic fluid pH that obtains in the step (2) is adjusted to 6.0, and adding papain (Nanning Pang Bo bioengineering Co., Ltd) 0.2g, at 55 ℃ of lower enzymolysis 3h, remove insoluble impurities by centrifugal, filtration, be prepared into laver fermentation enzymolysis liquid.
4, the preparation of laver proteoglycan nutriment powder: the laver fermentation enzymolysis liquid that obtains in the step (3) is prepared into laver proteoglycan nutriment powder by freeze drying.Recovery rate, main component, protein solubility and the amino acid of the laver proteoglycan nutriment powder that obtains formed analyze, the result is shown in table 1～4.
Embodiment 2
1, the preparation of bafillus natto bacterium liquid: the bafillus natto bacterial classification is provided by Microbiological Lab of bioengineering institute of Collects The American University, utilize the LB solid medium at 37 ℃ of lower 12h of cultivation, recycling LB fluid nutrient medium was at 37 ℃ of lower 24h of cultivation, for following laver fermentation after picking went out single bacterium colony.
2, the preparation of laver zymotic fluid: get the 8g laver and put into 200ml distilled water, add simultaneously the bacillus subtilis bacterium liquid 8ml of preparation in the step (1), at 30 ℃ of shaker fermentation 24h, be prepared into the laver zymotic fluid.
3, the enzymolysis of laver zymotic fluid: utilize acetic acid that the laver zymotic fluid pH that obtains in the step (2) is adjusted to 5.5, and adding papain (Nanning Pang Bo bioengineering Co., Ltd) 0.2g, at 55 ℃ of lower enzymolysis 2h, remove insoluble impurities by centrifugal, filtration, be prepared into laver fermentation enzymolysis liquid.
4, the preparation of laver proteoglycan nutriment powder: the preparation method is identical with embodiment 1.And recovery rate, main component, protein solubility and the amino acid of the laver proteoglycan nutriment powder that obtains formed analyze, the result is shown in table 1-4.
Table 1 has shown the result of the recovery rate of laver proteoglycan nutriment powder under the optimum condition, as seen from table, utilizes fermentation of bacillus subtilis again through papain enzymolysis, and laver protein extracting ratio, laver amylose recovery rate and solid content recovery rate all are the highest.But no matter utilize bacillus subtilis or utilize bafillus natto, its protein extracting ratio and polysaccharide extract rate all far above the laver amylose recovery rate (35%) that utilizes the ultrasonic disruption extraction method of present report and laver protein extracting ratio (45%) (Xiao Haifang. pulse ultrasonic wave extracts the technical study of albumen and polysaccharide in the laver simultaneously. Food Science, 2007,6).Main component to the laver proteoglycan nutriment powder of extraction among the embodiment 1,2 is measured, and the result is as shown in table 2.With respect to dried laver, utilize in the extract of fermentation of bacillus subtilis, protein content and polyoses content are basically identical, but utilize the polyoses content of extract of bacillus natto to ferment obviously lower, this might sweat in the Bacillus natto consumption than the used up cause of many parts glycocalix.Then we have also measured laver fermenting enzyme hydrolysis products protein solubility (table 3) under different pH values.As seen from table, bacillus subtilis and bafillus natto are not remarkable on the protein solubility impact of laver fermentation zymolyte.In addition, except the albumen solubility of two kinds of extracts under pH5 only has about 93%, under other pH all greater than 95%, shown that the laver proteoglycan nutriment powder that utilizes the present invention to extract has good protein dissolution performance, for the laver extract provides convenience as the natural additive Applications in food.Then, the amino acid composition of laver and extract thereof is analyzed, the result is as shown in table 4.As shown in Table 4, the amino acid contained total class of porphyra haitanensis is complete, and essential amino acid accounts for 47% of total amino acid content, has illustrated that laver albumen belongs to high-quality protein.And in the laver extract total amino content and must all not have to occur significantly to change by amino acid content, shown laver in the enzymolysis process that ferments, be that cell membrane is destroyed, protein is become at last solable matter and is dissolved in the water by enzymolysis.Therefore the laver proteoglycan nutriment powder that utilizes the present invention to produce has kept the original health nutrient composition of laver.
In sum, the present invention adds 1%～4% bacillus subtilis or bafillus natto bacterium liquid in a water purification bubble good dried laver, behind 30 ℃ of lower shaker fermentation 12～36h, recycling 0.05%～0.2% papain carries out enzymolysis 2～4h to the laver zymotic fluid, by steps such as heat sterilization, the enzyme that goes out, centrifugation, filtration, freeze drying or spray-dryings, can be prepared into the solubility nutrient powder that is rich in laver albumen and polysaccharide at last.The laver nutritive element powder of the present invention's preparation has the health nutrient composition same with laver, can be used as natural additive for foodstuff and is applied in food service industry, also can be used as intermediate raw material and researches and develops various novel functional foods.
Above-mentioned only is specific embodiments of the invention, but design concept of the present invention is not limited to this, allly utilizes this design that the present invention is carried out the change of unsubstantiality, all should belong to the behavior of invading protection domain of the present invention.
The recovery rate of table 1 laver proteoglycan nutriment powder
The main component of table 2 laver and extract thereof
The protein solubility (%) of table 3 laver extract
The amino acid of table 4 laver and extract thereof forms
Annotate: * represents essential amino acid.
Claims (8)
1. the production method of a main laver proteoglycan nutriment powder is characterized in that may further comprise the steps:
1) preparation laver zymotic fluid: add microbial inoculum in the laver aqueous solution that immersion is opened, shaker fermentation gets the laver zymotic fluid; Described microbial bacteria is bacillus subtilis or bafillus natto;
2) enzymolysis of laver zymotic fluid: in adjusting the laver zymotic fluid of pH, add papain enzymolysis, remove insoluble substance by heat sterilization, the enzyme that goes out, centrifugation, filtration, obtain laver fermentation enzymolysis liquid;
3) preparation of laver proteoglycan nutriment powder: with the laver enzymolysis liquid drying of fermenting, namely get laver proteoglycan nutriment powder.
2. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1, it is characterized in that in step 1), the concrete grammar of described preparation laver zymotic fluid is: first microbial bacteria is utilized the LB solid medium to cultivate 8～12h under 37 ° of C, picking is cultivated 16～24h with the LB fluid nutrient medium after going out single bacterium colony under 37 ° of C, microbial inoculum as fermentation usefulness, the recycling microbial inoculum carries out shaker fermentation to laver, is prepared into the laver zymotic fluid.
3. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1, it is characterized in that in step 1), the temperature of described shaker fermentation is 30 ° of C, and the time of described shaker fermentation is 12～36h, and the rotating speed of described shaker fermentation is 120～200rpm.
4. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1 is characterized in that in step 1), and the addition of described microbial inoculum is for soaking 1%～4% of the laver aqueous solution volume open.
5. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1 is characterized in that in step 2) in, described pH is 5.5～6.5.
6. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1 is characterized in that in step 2) in, the addition of described protease is 0.05%～0.2% of laver zymotic fluid by volume.
7. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1 is characterized in that in step 2) in, the temperature of described enzymolysis is 55 ° of C, the time of described enzymolysis is 2～4h.
8. the production method of a main laver proteoglycan nutriment powder as claimed in claim 1 is characterized in that in step 3), described dry freeze drying or the spray-drying of adopting.
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| CN104710541A (en) * | 2015-03-31 | 2015-06-17 | 广西还珠海洋生物科技有限公司 | Method for preparing laminarin from kelp |
| CN104705709B (en) * | 2015-04-07 | 2017-10-24 | 青岛海洋生物医药研究院股份有限公司 | A kind of method of pair of phase fermenting and producing marine alga ferment |
| CN105167011A (en) * | 2015-08-17 | 2015-12-23 | 济南舜昊生物科技有限公司 | Laver salad and preparation method therefor |
| CN106418097A (en) * | 2016-09-22 | 2017-02-22 | 辽宁天润生物技术有限公司 | Preparation method of soluble natto surface fermented product |
| CN106722423A (en) * | 2016-12-01 | 2017-05-31 | 浙江百惠生物科技有限公司 | A kind of seaweed ferment of seasoning |
| CN109369819A (en) * | 2018-10-22 | 2019-02-22 | 温州科技职业学院 | A kind of extracting method of Hijiki polysaccharide |
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