CN102212589B - Method for preparing bacterial cellulose - Google Patents
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- CN102212589B CN102212589B CN 201110110794 CN201110110794A CN102212589B CN 102212589 B CN102212589 B CN 102212589B CN 201110110794 CN201110110794 CN 201110110794 CN 201110110794 A CN201110110794 A CN 201110110794A CN 102212589 B CN102212589 B CN 102212589B
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Abstract
The invention relates to a method for preparing bacterial cellulose, in particular to a method for preparing bacterial cellulose by adding corn cob enzymolysis solution in a culture medium. The corn cob enzymolysis solution is prepared from corn cobs by enzymolysis of cellulase, hemi-cellulase and xylanase; and the corn cob enzymolysis solution is added into the culture medium for preparing the bacterial cellulose. By using common agricultural wastes, the production cost is reduced, and the regionalism of bacterial cellulose production is broken; and rich xylan in the corn cobs is subjected to enzymolysis to form xylooligosaccharide with good health-care function, and the xylooligosaccharide is wrapped or adsorbed in the bacterial cellulose, so that the health-care function of the bacterial cellulose is improved.
Description
Technical field
The present invention relates to the biological fermentation field, be specifically related to a kind of preparation method of bacteria cellulose, more specifically relate to a kind of method that the corn cob enzymolysis solution prepares bacteria cellulose of adding in substratum.
Background technology
Corn is one of China's Three major grain crops.According to statistics, annual production is about 1.2 hundred million tons.Can obtain simultaneously 1/3 kg corn core calculating by producing 1 kg corn, approximately 0.4 hundred million tons of China's corn cob annual production, output is extremely considerable.At present China to the utilization of corn cob comparatively speaking seldom, only has ten thousand tons of 40-50 to be used for furfural, wood sugar, Xylitol and phenols production, and the overwhelming majority is burnt as agricultural wastes.But, contain various saccharides in corn cob, especially contain a large amount of xylans, its Partial digestion just can obtain xylo-oligosaccharide, and xylo-oligosaccharide can make probiotic bacterium breed in a large number in human intestinal, has good nourishing function.
Bacteria cellulose (Bacterial cellulose, BC) refer under different condition, by synthetic cellulosic general designation such as certain microorganism in acetic acid Pseudomonas (Acetobacter), Agrobacterium (Agrobacterium), rhizobium (Rhizobium) and Sarcina (Sarcina) etc.The natural cellulose that bacteria cellulose and plant or marine alga produce has identical molecular structure unit, but bacteria cellulose fibre has the character of many uniquenesses.Such as high-crystallinity, high retentiveness, high biophase content and high biodegradability etc.The every field such as food, medical material, papermaking have been widely used in.
The production of bacteria cellulose is that the bacterial classification that will produce bacteria cellulose is inoculated in substratum, cultivates under certain condition, and the metabolism by microorganism produces bacteria cellulose film.Need to provide the required carbon source of growth, metabolism, nitrogenous source etc. in substratum.The used medium of bacteria cellulose production at present is mainly as main raw material with Sucus Cocois, can also add other fruit juice, as pineapple juice, various carbohydrates, as glucose, sucrose etc., the inorganic mineral such as the nitrogenous source such as peptone, extractum carnis, yeast extract and potassium primary phosphate, sal epsom.But the cost of carbohydrate is higher, and uses the tropical fruit fruit juice such as Sucus Cocois, has significant region, and the southern area that makes production only be adapted at abounding with tropical fruit carries out.Have no at present and add the corn cob enzymolysis solution in culture medium of bacterial cellulose report.
Summary of the invention
The object of the invention is to: a kind of preparation method of bacteria cellulose is provided, and the present invention has reduced production cost, has broken the region that bacteria cellulose is produced; And also contain functional low polyxylose in the corn cob enzymolysis solution, it can be wrapped or be adsorbed in the reticulated structure of bacteria cellulose film, when bacteria cellulose film is processed to food raw material when edible for the human or animal, it just can make the probiotic bacterium in enteron aisle breed in a large number, improves the nourishing function of bacteria cellulose product.
For achieving the above object, technical scheme of the present invention is: a kind of preparation method of bacteria cellulose is provided, comprises preparation substratum, inoculating strain, static fermentation, collection mycoderm and cleaning step, wherein, during the preparation substratum, added the corn cob enzymolysis solution.
Preferably, substratum is that corn cob enzymolysis solution and Sucus Cocois or distilled water were mixed by weight 1: 0.5～1: 2, then adds other carbon sources that account for substratum 10～30% weight, accounts for that the nitrogenous source of substratum 1～5% weight and micro-inorganic salt make.
Preferably, described corn cob enzymolysis solution is prepared by the method for following steps: corn cob is broken, weight ratio by corn cob and water added water in 1: 10～1: 20,100～120 ℃ of boilings 3～5 hours, obtain slurries, cooling rear with behind dilute hydrochloric acid adjust pH to 4.5～6.5, add cellulase, hemicellulase and zytase, add-on is respectively slurry weight: 0.1～0.3%, 0.05～0.1% and 0.05～0.1%, be incubated 1～2 hour under 50～60 ℃, intensification is gone out to filter after enzyme and is carried out solid-liquid separation, and filtrate is the corn cob enzymolysis solution.
Preferably, the weight ratio of corn cob and water is 1: 12.
Preferably, corn cob was in 120 ℃ of boilings 5 hours.
Preferably, add and the pH value is transferred to 6 before enzyme.
Preferably, the add-on of cellulase, hemicellulase and zytase is 0.3%, 0.1% and 0.1% of slurry weight.
Preferably, 55 ℃ are incubated enzymolysis 2 hours.
Preferably, substratum is that the corn enzymolysis solution is mixed by the weight ratio of 1: 1 with Sucus Cocois, then adds other carbon sources that account for substratum 15% weight, the nitrogenous source that accounts for substratum 2% weight and micro-inorganic salt to make; Wherein other carbon sources are one or more in glucose, sucrose; Nitrogenous source is one or more in peptone, extractum carnis, yeast extract; Inorganic salt are one or more in potassium primary phosphate or sal epsom.
The present invention also provides by the prepared bacteria cellulose product of aforesaid method.
The present invention compared with prior art, the present invention utilizes the agricultural by-products corn cob as the composition in culture medium of bacterial cellulose, reduced production cost, has broken the region that bacteria cellulose is produced; And also contain functional low polyxylose in the corn cob enzymolysis solution, it can be wrapped or be adsorbed in the reticulated structure of bacteria cellulose film, when bacteria cellulose film is processed to food raw material when edible for the human or animal, it just can make the probiotic bacterium in enteron aisle breed in a large number, improves the nourishing function of bacteria cellulose product.
Embodiment
The invention will be further described below in conjunction with preferred embodiment, but the present invention never is limited to following embodiment.
Embodiment 1:
Corn cob is broken, weight ratio by corn cob and water adds water at 1: 10,120 ℃ of boilings 5 hours obtain slurries, and are cooling rear with after dilute hydrochloric acid adjust pH to 4.5, add cellulase, hemicellulase and zytase, add-on is respectively slurry weight: 0.3%, 0.1% and 0.05%, and insulation is 2 hours under 50 ℃, then the enzyme that goes out that heats up, solid-liquid separation is carried out in filtration, and filtrate is the corn cob enzymolysis solution; The corn cob enzymolysis solution of getting 50ml mixes with the Sucus Cocois of 50ml, add again the potassium primary phosphate of glucose, 5g peptone and the trace of 30g to make substratum, access accounts for the gluconate pyracetobacillus seed liquor of substratum 12% weight, cultivated 15 days under 28 ℃, take out the bacteria cellulose film on liquid level, repeatedly clean to neutrality with deionized water.
Embodiment 2:
Corn cob is broken, weight ratio by corn cob and water adds water at 1: 15,110 ℃ of boilings 4 hours obtain slurries, and are cooling rear with after dilute hydrochloric acid adjust pH to 6, add cellulase, hemicellulase and zytase, add-on is respectively slurry weight: 0.2%, 0.08% and 0.1%, and insulation is 1 hour under 60 ℃, then the enzyme that goes out that heats up, solid-liquid separation is carried out in filtration, and filtrate is the corn cob enzymolysis solution.The corn cob enzymolysis solution of getting 50ml mixes with the distilled water of 50ml, add again the potassium primary phosphate of glucose, 1g peptone and the trace of 10g to make substratum, access accounts for the gluconate pyracetobacillus seed liquor of substratum 10% weight, cultivated 8 days under 30 ℃, take out the bacteria cellulose film on liquid level, repeatedly clean to neutrality with deionized water.
Embodiment 3:
Corn cob is broken, weight ratio by corn cob and water adds water at 1: 20,100 ℃ of boilings 5 hours obtain slurries, and are cooling rear with after dilute hydrochloric acid adjust pH to 6.5, add cellulase, hemicellulase and zytase, add-on is respectively slurry weight: 0.1%, 0.05% and 0.08%, and insulation is 2 hours under 55 ℃, then the enzyme that goes out that heats up, solid-liquid separation is carried out in filtration, and filtrate is the corn cob enzymolysis solution.The corn cob enzymolysis solution of getting 50ml mixes with the Sucus Cocois of 50ml, add again the potassium primary phosphate of glucose, 3g peptone and the trace of 20g to make substratum, access accounts for the gluconate pyracetobacillus seed liquor of substratum 8% weight, cultivated 12 days under 29 ℃, take out the bacteria cellulose film on liquid level, repeatedly clean to neutrality with deionized water.
Reference examples:
Get glucose 50g, peptone 5g, potassium primary phosphate 2g mixing, complement to 100ml with distilled water, be mixed with fermentation medium for bacterial cellulose, the scheme according to above-described embodiment makes bacteria cellulose film afterwards.
Experimental example: the sensory evaluation of bacteria cellulose film
Set up 5 people's expertise groups, the setting assessment item is: color and luster, mouthfeel, transparency; Evaluation method is: color and luster: oyster white gets full marks, and color is crossed dark or crossed shallow all corresponding subtract minute, score value 1～5 minute; Mouthfeel: mouthfeel is moderate to get full marks, really up to the mark or cross soft all corresponding subtract minute, score value 1～5 minute; Transparency: translucent getting full marks, transparent or opaque all corresponding subtract minute, score value 1～5 minute.Each assessment item is got 5 experts' mean value as estimating score.Evaluation result sees the following form:
Table 1: the Analyses Methods for Sensory Evaluation Results of bacteria cellulose film
| Evaluation result | Color and luster | Mouthfeel | Transparency |
| Embodiment 1 | 5 | 5 | 5 |
| Embodiment 2 | 5 | 4 | 4 |
| Embodiment 3 | 4 | 4 | 5 |
| Reference examples | 4 | 5 | 5 |
Aesthetic quality and the no significant difference of bacteria cellulose film when when as seen adopting the corn cob enzymolysis solution as carbon source, the aesthetic quality of bacteria cellulose film and direct use glucose are as carbon source.
Above disclosed is only preferred embodiment of the present invention, certainly can not limit with this interest field of the present invention, and the equivalent variations of therefore doing according to claim of the present invention still belongs to the scope that the present invention is contained.
Claims (8)
1. the preparation method of a bacteria cellulose, comprise preparation substratum, inoculating strain, static fermentation, collection mycoderm and cleaning step, it is characterized in that: during the preparation substratum, added the corn cob enzymolysis solution; Wherein said corn cob enzymolysis solution is prepared by the method for following steps: corn cob is broken, weight ratio by corn cob and water added water in 1: 10～1: 20,100～120 ℃ of boilings 3～5 hours, obtain slurries, cooling rear with behind dilute hydrochloric acid adjust pH to 4.5～6.5, add cellulase, hemicellulase and zytase, add-on is respectively slurry weight: 0.1～0.3%, 0.05～0.1% and 0.05～0.1%, be incubated 1～2 hour under 50～60 ℃, intensification is gone out to filter after enzyme and is carried out solid-liquid separation, and filtrate is the corn cob enzymolysis solution.
2. the preparation method of bacteria cellulose according to claim 1, it is characterized in that: substratum is that the corn cob enzymolysis solution is mixed by weight 1: 0.5～1: 2 with Sucus Cocois or distilled water, add again other carbon sources that account for substratum 10～30% weight, account for that the nitrogenous source of substratum 1～5% weight and micro-inorganic salt make.
3. the preparation method of bacteria cellulose according to claim 1, it is characterized in that: the weight ratio of corn cob and water is 1: 12.
4. the preparation method of bacteria cellulose according to claim 1, it is characterized in that: corn cob was in 120 ℃ of boilings 5 hours.
5. the preparation method of bacteria cellulose according to claim 1, is characterized in that: before adding enzyme, the pH value is transferred to 6.
6. the preparation method of bacteria cellulose according to claim 1, it is characterized in that: the add-on of cellulase, hemicellulase and zytase is 0.3%, 0.1% and 0.1% of slurry weight.
7. the preparation method of bacteria cellulose according to claim 1 is characterized in that: 55 ℃ of insulation enzymolysis 2 hours.
8. the preparation method of bacteria cellulose according to claim 2, it is characterized in that: substratum is that the corn enzymolysis solution is mixed by the weight ratio of 1: 1 with Sucus Cocois, then adds other carbon sources that account for substratum 15% weight, the nitrogenous source that accounts for substratum 2% weight and micro-inorganic salt to make; Wherein other carbon sources are one or more in glucose, sucrose; Nitrogenous source is one or more in peptone, extractum carnis, yeast extract; Inorganic salt are one or more in potassium primary phosphate or sal epsom.
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| CN102690773B (en) * | 2012-06-18 | 2014-01-01 | 南开大学 | Enterobacteria strain FY-07 and method thereof for producing bacterial cellulose by static liquid submerged fermentation |
| CN105420310A (en) * | 2015-12-16 | 2016-03-23 | 贵州大学 | Method for improving yield and quality of bacterial cellulose dry film by optimizing rice saccharification liquid fermentation culture medium |
| CN105861622B (en) * | 2016-05-26 | 2019-07-09 | 天津科技大学 | A kind of viable count method of bacteria cellulose production bacterial strain |
| CN106099182B (en) * | 2016-08-05 | 2018-08-03 | 宁波高智科技咨询服务有限公司 | A kind of lithium battery bacterial cellulose gel method for preparing polymer electrolytes |
| CN107698805A (en) * | 2016-08-09 | 2018-02-16 | 丁玉琴 | A kind of defervescence plaster used preparation method with compound water congealing glued membrane |
| CN107586801A (en) * | 2017-10-19 | 2018-01-16 | 南京理工大学 | A kind of method that bacteria cellulose is prepared using cotton stalk |
| CN108642106A (en) * | 2018-05-04 | 2018-10-12 | 天津科技大学 | A kind of method of a large amount of acquisition wood gluconic acid acetobacter thalline |
| CN109468341A (en) * | 2018-10-07 | 2019-03-15 | 南宁润生堂生物科技有限公司 | A kind of production method of fermented tea mycoderm zymolyte |
| CN111484901B (en) * | 2019-01-28 | 2021-09-28 | 海南光宇生物科技有限公司 | Enzyme-containing laundry detergent and preparation method thereof |
| CN111484902B (en) * | 2019-01-28 | 2021-09-28 | 海南光宇生物科技有限公司 | Preparation method of concentrated enzyme-containing laundry beads |
| CN112515099A (en) * | 2019-09-18 | 2021-03-19 | 钟春燕 | Method for processing instant noodles at low temperature |
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| CN101979636B (en) * | 2010-11-05 | 2013-03-20 | 钟春燕 | Method for preparing spherical bacterial cellulose |
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