CN107164428A - A kind of method for preparing bacteria cellulose as carbon source with ligocellulose degradation's liquid - Google Patents
A kind of method for preparing bacteria cellulose as carbon source with ligocellulose degradation's liquid Download PDFInfo
- Publication number
- CN107164428A CN107164428A CN201710595452.2A CN201710595452A CN107164428A CN 107164428 A CN107164428 A CN 107164428A CN 201710595452 A CN201710595452 A CN 201710595452A CN 107164428 A CN107164428 A CN 107164428A
- Authority
- CN
- China
- Prior art keywords
- bacterial cellulose
- lignocellulose
- degradation
- liquid
- deionized water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000007788 liquid Substances 0.000 title claims abstract description 22
- 230000015556 catabolic process Effects 0.000 title claims abstract description 20
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 20
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 19
- 238000000034 method Methods 0.000 title claims abstract description 16
- 229910052799 carbon Inorganic materials 0.000 title claims abstract description 11
- 229920002678 cellulose Polymers 0.000 title claims description 16
- 239000001913 cellulose Substances 0.000 title claims description 16
- 241000894006 Bacteria Species 0.000 title claims description 8
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 54
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000012528 membrane Substances 0.000 claims abstract description 17
- 239000008367 deionised water Substances 0.000 claims abstract description 16
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 16
- 238000000855 fermentation Methods 0.000 claims abstract description 13
- 230000004151 fermentation Effects 0.000 claims abstract description 13
- 241000209140 Triticum Species 0.000 claims abstract description 10
- 235000021307 Triticum Nutrition 0.000 claims abstract description 10
- 239000001963 growth medium Substances 0.000 claims abstract description 5
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 108010059892 Cellulase Proteins 0.000 claims abstract description 4
- 229940106157 cellulase Drugs 0.000 claims abstract description 4
- 239000002609 medium Substances 0.000 claims description 12
- 239000001888 Peptone Substances 0.000 claims description 9
- 108010080698 Peptones Proteins 0.000 claims description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 9
- 235000019319 peptone Nutrition 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 5
- 238000001784 detoxification Methods 0.000 claims 3
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims 1
- 235000009754 Vitis X bourquina Nutrition 0.000 claims 1
- 235000012333 Vitis X labruscana Nutrition 0.000 claims 1
- 240000006365 Vitis vinifera Species 0.000 claims 1
- 235000014787 Vitis vinifera Nutrition 0.000 claims 1
- 239000012978 lignocellulosic material Substances 0.000 claims 1
- 238000006213 oxygenation reaction Methods 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 239000002023 wood Substances 0.000 claims 1
- 229920002749 Bacterial cellulose Polymers 0.000 abstract description 21
- 239000005016 bacterial cellulose Substances 0.000 abstract description 21
- 239000010902 straw Substances 0.000 abstract description 12
- 235000015097 nutrients Nutrition 0.000 abstract description 6
- 230000007071 enzymatic hydrolysis Effects 0.000 abstract description 5
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 abstract description 5
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 4
- 241000032681 Gluconacetobacter Species 0.000 abstract description 3
- 239000012620 biological material Substances 0.000 abstract description 3
- 238000011081 inoculation Methods 0.000 abstract description 3
- 239000006228 supernatant Substances 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 2
- 239000002699 waste material Substances 0.000 abstract description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical group OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 15
- 239000008103 glucose Substances 0.000 description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 239000000843 powder Substances 0.000 description 8
- 230000007935 neutral effect Effects 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- 229920002307 Dextran Polymers 0.000 description 6
- 238000002791 soaking Methods 0.000 description 5
- 244000235858 Acetobacter xylinum Species 0.000 description 3
- 235000002837 Acetobacter xylinum Nutrition 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 238000004042 decolorization Methods 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000013028 medium composition Substances 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000010903 husk Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开一种由木质纤维素降解液作碳源制备细菌纤维素的方法，属于生物材料降解及制备技术领域，包括下列步骤：首先进行木质纤维素原料预处理。然后利用纤维素酶降解处理后的木质纤维素。酶解后的降解液经过脱毒，然后离心、过滤，得上清液，利用HPLC测定其中酶解所产生的可发酵糖浓度后，添加其他营养物质配制成木葡糖酸醋杆菌的培养基，接种后，放入30℃的恒温培养箱中培养7d，产生细菌纤维素膜。取出膜后用去离子水和氢氧化钠溶液清理干净，得到纯净的细菌纤维素膜。本发明生产的细菌纤维素膜具有较好的持水性，生物相容性和较高的力学强度。本发明中木质纤维素原料为小麦秸秆，其来源广泛，对于减少资源浪费，保护环境，降低发酵成本，具有重要意义。
The invention discloses a method for preparing bacterial cellulose by using lignocellulose degradation liquid as a carbon source, which belongs to the technical field of biomaterial degradation and preparation, and comprises the following steps: firstly, lignocellulose raw material is pretreated. The treated lignocellulose is then degraded by cellulase. The degradation solution after enzymatic hydrolysis is detoxified, then centrifuged and filtered to obtain the supernatant, and the concentration of fermentable sugar produced by enzymatic hydrolysis is measured by HPLC, and other nutrients are added to prepare the culture medium of Gluconacetobacter xylinum , after inoculation, placed in a constant temperature incubator at 30°C for 7 days to produce bacterial cellulose membranes. After taking out the membrane, clean it with deionized water and sodium hydroxide solution to obtain a pure bacterial cellulose membrane. The bacterial cellulose membrane produced by the invention has better water holding capacity, biocompatibility and higher mechanical strength. In the present invention, the lignocellulose raw material is wheat straw, which has a wide range of sources, and is of great significance for reducing waste of resources, protecting the environment, and reducing fermentation costs.
Description
技术领域technical field
本发明涉及生物材料的降解及其制备技术领域，特别涉及一种以木质纤维素降解液作碳源制备细菌纤维素的方法。The invention relates to the technical field of biomaterial degradation and preparation thereof, in particular to a method for preparing bacterial cellulose by using lignocellulose degradation liquid as a carbon source.
背景技术Background technique
细菌纤维素作为一种新型的生物材料，具有优良的物理和化学性能：高纯度，高持水性，透气性，高抗张强度和弹性，良好的生物亲和性，生物相容性。使其在医疗，食品，造纸，化妆品等多个行业领域具有广泛的应用前景。As a new type of biological material, bacterial cellulose has excellent physical and chemical properties: high purity, high water holding capacity, air permeability, high tensile strength and elasticity, good bio-affinity, and biocompatibility. It has broad application prospects in many industries such as medical treatment, food, papermaking, cosmetics and so on.
木质纤维素是地球上最丰富的可再生资源，每年仅陆生植物就可以产生木质纤维素大约500亿吨，占地球生物总量的60-80％。我国拥有非常丰富的木质纤维素原料，仅仅农作物秸秆、皮壳一项，每年产量就达到4亿多吨，其中玉米秸杆(35％)，小麦秸杆(21％)和水稻秸秆(19％)是我国的三大秸秆。农作物秸秆主要以焚烧和粉碎的方式处理，不仅造成资源的浪费，而且污染环境。Lignocellulose is the most abundant renewable resource on the earth, and terrestrial plants alone can produce about 50 billion tons of lignocellulose every year, accounting for 60-80% of the total biomass on the earth. my country has very rich lignocellulosic raw materials, and the annual output of crop straw and husk alone reaches more than 400 million tons, of which corn straw (35%), wheat straw (21%) and rice straw (19%) ) are the three major straws in our country. Crop stalks are mainly disposed of by incineration and crushing, which not only wastes resources but also pollutes the environment.
细菌纤维素由吡喃型葡萄糖残基通过β-1，4-糖苷键连接而成，木质纤维素是D-葡萄糖以β-1，4-糖苷键组成的大分子多糖，二者具有同源性。将木质纤维素降解，提供碳源进行细菌纤维素的合成是一种有效的木质纤维素处理方式，尤其在世界人口不断增长，资源紧缺和环境污染日益严重的情况下，利用木质纤维素降解合成细菌纤维素具有划时代意义和广泛的应用前景。Bacterial cellulose is composed of glucopyranose residues connected by β-1,4-glycosidic bonds, and lignocellulose is a macromolecular polysaccharide composed of D-glucose with β-1,4-glycosidic bonds. The two have homology sex. Degrading lignocellulose and providing carbon sources for the synthesis of bacterial cellulose is an effective way to treat lignocellulose, especially in the context of growing world population, shortage of resources and increasingly serious environmental pollution, using lignocellulose to degrade and synthesize bacteria Bacterial cellulose has epoch-making significance and broad application prospects.
发明内容Contents of the invention
本发明涉及提供一种以木质纤维素降解液作碳源制备细菌纤维素的方法，该方法绿色环保，简单易行，既解决了环境压力又创造了经济价值，为其他行业提供了一种优质材料。The invention relates to providing a method for preparing bacterial cellulose by using lignocellulose degradation liquid as a carbon source. The method is environmentally friendly, simple and easy to implement, solves environmental pressure and creates economic value, and provides a high-quality cellulose for other industries. Material.
为解决上述技术问题，本发明采用的技术方案为：In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
一种以木质纤维素降解液作碳源制备细菌纤维素的方法，包括下列步骤：A method for preparing bacterial cellulose with lignocellulose degradation liquid as carbon source, comprising the following steps:
(1)将剪成小段的小麦秸秆与0.5M NaOH溶液按料液比1∶20混合，于高压蒸汽灭菌锅中121℃预处理1小时，预处理后的样品经过纱布过滤，并用大量的去离子水洗涤至洗液呈中性。弃去上清液，保留固体残渣，待晾干后粉碎并过20目筛。(1) Mix the wheat straw cut into small pieces with 0.5M NaOH solution at a material-to-liquid ratio of 1:20, pretreat it in a high-pressure steam sterilizer at 121°C for 1 hour, filter the pretreated sample through gauze, and wash it with a large amount of Wash with deionized water until the washing liquid is neutral. Discard the supernatant, keep the solid residue, pulverize and pass through a 20-mesh sieve after drying.
(2)酶解反应的总体积为200mL，在1000mL的三角瓶中进行，葡聚糖负荷为4％，加入纤维素降解混合酶液，纤维素酶按20FPU/g葡聚糖的负荷添加，然后180rpm，50℃摇床降解。其中葡聚糖负荷定义为葡聚糖的质量在总的固液混合物体积中的百分比(认为小麦秸秆的密度大约为1g/ml)。(2) The total volume of the enzymolysis reaction is 200mL, carried out in a 1000mL Erlenmeyer flask, the load of dextran is 4%, the mixed enzyme solution for cellulose degradation is added, and the cellulase is added at a load of 20FPU/g dextran, Then 180rpm, 50°C shaker degradation. Wherein the dextran load is defined as the percentage of the mass of dextran in the total volume of the solid-liquid mixture (the density of wheat straw is considered to be about 1 g/ml).
(3)酶解液经离心，过滤，除去固体残渣后进行脱毒。先用饱和Ca(OH)2调节pH至10.0，30℃，静置10h，然后加入6.0mol/L H2SO4调节pH至5.0，加入质量分数5％的活性炭，100rpm，40℃，振荡3h。然后过滤进行高温灭菌得到水解液。最后配制发酵培养基，测定酶解液的葡萄糖浓度，然后稀释到相应的浓度，添加除碳源外的其他营养物质。发酵液培养基成分(g/L)：葡萄糖15-25，蛋白胨6-10，酵母粉4.5-7.5，Na2HPO3 6-10。pH为6.0。(3) The enzymolysis solution is centrifuged, filtered, and detoxified after removing solid residues. First use saturated Ca(OH) 2 to adjust the pH to 10.0, 30°C, let stand for 10h, then add 6.0mol/L H 2 SO 4 to adjust the pH to 5.0, add 5% activated carbon, 100rpm, 40°C, shake for 3h. Then filter and sterilize at high temperature to obtain a hydrolyzate. Finally, the fermentation medium is prepared, the glucose concentration of the enzymolysis solution is measured, and then diluted to the corresponding concentration, and other nutrients except the carbon source are added. Fermentation medium composition (g/L): glucose 15-25, peptone 6-10, yeast powder 4.5-7.5, Na 2 HPO 3 6-10. The pH is 6.0.
(4)先活化木葡糖酸醋杆菌，然后按照体积比3％的接种量把种子液接种到发酵液中，然后放入30℃的恒温培养箱中培养7d，发酵液表面形成细菌纤维素膜。(4) First activate Gluconoacetobacter xylinum, then inoculate the seed liquid into the fermentation broth according to the inoculation amount of 3% by volume, and then put it into a constant temperature incubator at 30°C for 7 days, and bacterial cellulose is formed on the surface of the fermentation broth membrane.
(5)取出纤维素膜用去离子水清洗干净，然后用0.1MNaOH溶液浸泡至纤维素膜呈白色，要多次更换NaOH溶液，最后用去离子水浸泡至浸泡液pH为中性。(5) Take out the cellulose membrane and clean it with deionized water, then soak it with 0.1M NaOH solution until the cellulose membrane is white, change the NaOH solution several times, and finally soak it with deionized water until the pH of the soaking solution is neutral.
优选的，本发明选择小麦秸秆作为木质纤维素原材料，是因为其简单易得，经过合适预处理后的小麦秸秆纤维素含量高，本发明保护范围不仅仅局限于此种植物。Preferably, the present invention selects wheat straw as the lignocellulose raw material because it is easy to obtain, and the wheat straw after proper pretreatment has high cellulose content, and the protection scope of the present invention is not limited to this kind of plant.
优选的，本发明选择酶降解木质纤维素，是因为其简单易行，降解效率高。Preferably, the present invention chooses enzyme to degrade lignocellulose because it is easy to implement and has high degradation efficiency.
本发明的有益效果：Beneficial effects of the present invention:
(1)本发明提供了一种以木质纤维素降解液作碳源制备细菌纤维素的方法，木质纤维素材料来源丰富，价格低廉，有大规模实际应用的前景。(1) The present invention provides a method for preparing bacterial cellulose by using lignocellulose degradation liquid as a carbon source. The source of lignocellulose material is abundant, the price is low, and there is a prospect of large-scale practical application.
(2)木质纤维素的有效利用既解决了环境压力，又创造了经济价值。(2) The effective utilization of lignocellulose not only solves the environmental pressure, but also creates economic value.
(3)生成的细菌纤维素，基于其高纯度，高持水性，透气性，高抗张强度和弹性，良好的生物亲和性，生物相容性，可广泛应用于造纸，食品，医药，纺织，航空航天等领域。(3) The resulting bacterial cellulose can be widely used in papermaking, food, medicine, Textile, aerospace and other fields.
附图说明Description of drawings
图1是小麦秸秆水解液产生的细菌纤维素的SEM图Figure 1 is the SEM image of bacterial cellulose produced from wheat straw hydrolyzate
图2是葡萄糖培养基产生的细菌纤维素的SEM图Fig. 2 is the SEM picture of the bacterial cellulose produced by glucose medium
具体实施方式detailed description
下面对本发明实施例中的技术方案进行清楚，完整的描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下获得的所有其他实施例，都属于本发明保护的范围。The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.
一种以木质纤维素降解液作碳源制备细菌纤维素的方法，包括以下步骤：A method for preparing bacterial cellulose with lignocellulose degradation liquid as carbon source, comprising the following steps:
(1)木质纤维素碱处理：将剪成小段的小麦秸秆与0.5M NaOH溶液按料液比1∶20混合，于高压蒸汽灭菌锅中121℃预处理1小时，预处理后的样品经过纱布过滤，并用大量的去离子水洗涤至洗液呈中性。弃去上清液，保留固体残渣，待晾干后粉碎并过20目筛。(1) Alkali treatment of lignocellulose: mix small pieces of wheat straw with 0.5M NaOH solution at a material-to-liquid ratio of 1:20, pretreat in a high-pressure steam sterilizer at 121°C for 1 hour, and pretreat the sample through Filter through gauze and wash with a large amount of deionized water until the washing liquid is neutral. Discard the supernatant, keep the solid residue, pulverize and pass through a 20-mesh sieve after drying.
(2)酶解：反应的总体积为200mL，在1000mL的三角瓶中进行，葡聚糖负荷为4％，加入纤维素降解混合酶液，纤维素酶按20FPU/g葡聚糖的负荷添加，然后180rpm，50℃摇床降解。(2) Enzymolysis: The total volume of the reaction is 200mL, carried out in a 1000mL Erlenmeyer flask, the load of dextran is 4%, the mixed enzyme solution for cellulose degradation is added, and the cellulase is added according to the load of 20FPU/g dextran , and then 180rpm, 50°C shaker degradation.
(3)酶解液经离心，过滤，除去固体残渣后进行脱毒。先用饱和Ca(OH)2调节pH至10.0，30℃，静置10h，然后加入6.0mol/L H2SO4调节pH至5.0，加入质量分数5％的活性炭，100rpm，40℃，振荡3h。然后过滤进行高温灭菌得到水解液。最后配制发酵培养基，测定酶解液的葡萄糖浓度，然后稀释到相应的浓度，添加除碳源外的其他营养物质。发酵液培养基成分(g/L)：葡萄糖15-25，蛋白胨6-10，酵母粉4.5-7.5，Na2HPO3 6-10。pH为6.0。(3) The enzymolysis solution is centrifuged, filtered, and detoxified after removing solid residues. First use saturated Ca(OH) 2 to adjust the pH to 10.0, 30°C, let stand for 10h, then add 6.0mol/L H 2 SO 4 to adjust the pH to 5.0, add 5% activated carbon, 100rpm, 40°C, shake for 3h. Then filter and sterilize at high temperature to obtain a hydrolyzate. Finally, the fermentation medium is prepared, the glucose concentration of the enzymolysis solution is measured, and then diluted to the corresponding concentration, and other nutrients except the carbon source are added. Fermentation medium composition (g/L): glucose 15-25, peptone 6-10, yeast powder 4.5-7.5, Na 2 HPO 3 6-10. The pH is 6.0.
(4)先活化木葡糖酸醋杆菌，然后按照体积比3％的接种量把种子液接种到发酵液中，然后放入30℃的恒温培养箱中培养7d，发酵液表面形成细菌纤维素膜。(4) First activate Gluconoacetobacter xylinum, then inoculate the seed liquid into the fermentation broth according to the inoculation amount of 3% by volume, and then put it into a constant temperature incubator at 30°C for 7 days, and bacterial cellulose is formed on the surface of the fermentation broth membrane.
(5)取出纤维素膜用去离子水清洗干净，然后用0.1M NaOH溶液浸泡至纤维素膜变白，要多次更换NaOH溶液，最后用去离子水浸泡至浸泡液pH为中性。(5) Take out the cellulose membrane and clean it with deionized water, then soak it with 0.1M NaOH solution until the cellulose membrane turns white, replace the NaOH solution several times, and finally soak it with deionized water until the pH of the soaking solution is neutral.
实施例1Example 1
(1)木质纤维素酶解之后，经过HPLC分析，葡萄糖浓度为30g/L。(1) After enzymatic hydrolysis of lignocellulose, through HPLC analysis, the glucose concentration is 30g/L.
(2)将降解液的葡萄糖浓度稀释到25g/L，添加其他营养物质，添加比例为：蛋白胨10g/L，酵母粉7.5g/L，Na2HPO3 10g/L，pH调节至6.0。在121℃下高压蒸汽灭菌20min。(2) Dilute the glucose concentration of the degradation solution to 25g/L, and add other nutrients at the following ratio: peptone 10g/L, yeast powder 7.5g/L, Na 2 HPO 3 10g/L, and adjust the pH to 6.0. Autoclaved at 121°C for 20min.
(3)把甘油管中保藏的木醋杆菌(Gluconacetobacter xylinus)CGMCC 2955接种到固体平板培养基上，放入30℃恒温培养箱中，培养2d后，得到活化菌。称取葡萄糖5.0g，蛋白胨2.0g，酵母粉1.5g，Na2HPO3 2.0g，pH为6.0，配制成200ml种子液培养基。将活化好的菌种接到种子液培养基溶液中30℃，180rpm振荡培养20h，得到种子液。(3) Inoculate the Gluconacetobacter xylinus CGMCC 2955 preserved in the glycerol tube onto the solid plate medium, put it into a constant temperature incubator at 30° C., and cultivate it for 2 days to obtain activated bacteria. Weigh 5.0 g of glucose, 2.0 g of peptone, 1.5 g of yeast powder, 2.0 g of Na2HPO 3 , pH 6.0, and prepare 200 ml of seed liquid culture medium. The activated strains were placed in the seed liquid medium solution at 30° C. and shaken at 180 rpm for 20 h to obtain the seed liquid.
(4)吸取6ml种子液接种到发酵培养基中，然后放入30℃的恒温培养箱中培养7d，发酵得到细菌纤维素膜。(4) Draw 6ml of seed solution and inoculate it into the fermentation medium, then put it into a constant temperature incubator at 30° C. and cultivate it for 7 days, and ferment to obtain bacterial cellulose membrane.
(5)取出纤维素膜用去离子水清洗干净，然后用0.1M NaOH溶液浸泡脱色，要多次更换NaOH溶液，最后用去离子水浸泡至浸泡液pH为中性。(5) Take out the cellulose membrane and clean it with deionized water, then soak it in 0.1M NaOH solution for decolorization, change the NaOH solution several times, and finally soak it with deionized water until the pH of the soaking solution is neutral.
实施例2Example 2
(1)木质纤维素酶解之后，经过HPLC分析，葡萄糖浓度为30g/L。(1) After enzymatic hydrolysis of lignocellulose, through HPLC analysis, the glucose concentration is 30g/L.
(2)将降解液的葡萄糖浓度稀释到20g/L，添加其他营养物质，添加比例为：蛋白胨8g/L，酵母粉6g/L，Na2HPO3 8g/L，pH调节至6.0。在121℃下高压蒸汽灭菌20min。(2) Dilute the glucose concentration of the degradation solution to 20g/L, and add other nutrients in the ratio: peptone 8g/L, yeast powder 6g/L, Na 2 HPO 3 8g/L, and adjust the pH to 6.0. Autoclaved at 121°C for 20min.
(3)把甘油管中保藏的木醋杆菌(Gluconacetobacter xylinus)CGMCC 2955接种到固体平板培养基上，放入30℃恒温培养箱中，培养2d后，得到活化菌。称取葡萄糖5.0g，蛋白胨2.0g，酵母粉1.5g，Na2HPO3 2.0g，pH为6.0，配制成200ml种子液培养基。将活化好的菌种接到种子液培养基溶液中30℃，180rpm振荡培养20h，得到种子液。(3) Inoculate the Gluconacetobacter xylinus CGMCC 2955 preserved in the glycerol tube onto the solid plate medium, put it into a constant temperature incubator at 30° C., and cultivate it for 2 days to obtain activated bacteria. Weigh 5.0 g of glucose, 2.0 g of peptone, 1.5 g of yeast powder, 2.0 g of Na2HPO 3 , pH 6.0, and prepare 200 ml of seed liquid culture medium. The activated strains were placed in the seed liquid medium solution at 30° C. and shaken at 180 rpm for 20 h to obtain the seed liquid.
(4)吸取6ml种子液接种到发酵培养基中，然后放入30℃的恒温培养箱中培养7d，发酵得到细菌纤维素膜。(4) Draw 6ml of seed solution and inoculate it into the fermentation medium, then put it into a constant temperature incubator at 30° C. and cultivate it for 7 days, and ferment to obtain bacterial cellulose membrane.
(5)取出纤维素膜用去离子水清洗干净，然后用0.1MNaOH溶液浸泡脱色，要多次更换NaOH溶液，最后用去离子水浸泡至浸泡液pH为中性。(5) Take out the cellulose membrane and clean it with deionized water, then soak it in 0.1M NaOH solution for decolorization, change the NaOH solution several times, and finally soak it with deionized water until the pH of the soaking solution is neutral.
实施例3Example 3
(1)木质纤维素酶解之后，经过HPLC分析，葡萄糖浓度为30g/L。(1) After enzymatic hydrolysis of lignocellulose, through HPLC analysis, the glucose concentration is 30g/L.
(2)将降解液的葡萄糖浓度稀释到15g/L，添加其他营养物质，添加比例为：蛋白胨6g/L，酵母粉7.5g/L，Na2HPO3 6g/L，pH调节至6.0。在121℃下高压蒸汽灭菌20min。(2) Dilute the glucose concentration of the degradation solution to 15g/L, and add other nutrients at the following ratio: peptone 6g/L, yeast powder 7.5g/L, Na 2 HPO 3 6g/L, and adjust the pH to 6.0. Autoclaved at 121°C for 20min.
(3)把甘油管中保藏的木醋杆菌(Gluconacetobacter xylinus)CGMCC 2955接种到固体平板培养基上，放入30℃恒温培养箱中，培养2d后，得到活化菌。称取葡萄糖5.0g，蛋白胨2.0g，酵母粉1.5g，Na2HPO3 2.0g，pH为6.0，配制成200ml种子液培养基。将活化好的菌种接到种子液培养基溶液中30℃，180rpm振荡培养20h，得到种子液。(3) Inoculate the Gluconacetobacter xylinus CGMCC 2955 preserved in the glycerol tube onto the solid plate medium, put it into a constant temperature incubator at 30° C., and cultivate it for 2 days to obtain activated bacteria. Weigh 5.0 g of glucose, 2.0 g of peptone, 1.5 g of yeast powder, 2.0 g of Na2HPO 3 , pH 6.0, and prepare 200 ml of seed liquid culture medium. The activated strains were placed in the seed liquid medium solution at 30° C. and shaken at 180 rpm for 20 h to obtain the seed liquid.
(4)吸取6ml种子液接种到发酵培养基中，然后放入30℃的恒温培养箱中培养7d，发酵得到细菌纤维素膜。(4) Draw 6ml of seed solution and inoculate it into the fermentation medium, then put it into a constant temperature incubator at 30° C. and cultivate it for 7 days, and ferment to obtain bacterial cellulose membrane.
(5)取出纤维素膜用去离子水清洗干净，然后用0.1M NaOH溶液浸泡脱色，要多次更换NaOH溶液，最后用去离子水浸泡至浸泡液pH为中性。(5) Take out the cellulose membrane and clean it with deionized water, then soak it in 0.1M NaOH solution for decolorization, change the NaOH solution several times, and finally soak it with deionized water until the pH of the soaking solution is neutral.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710595452.2A CN107164428A (en) | 2017-07-18 | 2017-07-18 | A kind of method for preparing bacteria cellulose as carbon source with ligocellulose degradation's liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710595452.2A CN107164428A (en) | 2017-07-18 | 2017-07-18 | A kind of method for preparing bacteria cellulose as carbon source with ligocellulose degradation's liquid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107164428A true CN107164428A (en) | 2017-09-15 |
Family
ID=59817939
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710595452.2A Pending CN107164428A (en) | 2017-07-18 | 2017-07-18 | A kind of method for preparing bacteria cellulose as carbon source with ligocellulose degradation's liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107164428A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108315371A (en) * | 2018-04-20 | 2018-07-24 | 北京理工大学珠海学院 | A kind of cultural method of bacteria cellulose |
| CN109022534A (en) * | 2018-08-01 | 2018-12-18 | 淮阴师范学院 | Utilize the method and corn stover degradation product fuel cell of corn stover production bacteria cellulose film |
| CN109182187A (en) * | 2018-09-20 | 2019-01-11 | 天津科技大学 | A method of utilizing glucose mass transfer in agar measurement Bacterial Cellulose in Fermentation Condition |
| CN110218751A (en) * | 2019-06-03 | 2019-09-10 | 广西大学 | The method for preparing cellobiose and cellooligosaccharide using carbohydrate |
| CN113234637A (en) * | 2021-06-16 | 2021-08-10 | 南开大学 | Fermentation medium for large-scale efficient production of bacterial cellulose and fermentation method thereof |
| CN113913971A (en) * | 2020-07-10 | 2022-01-11 | 南京理工大学 | A kind of method for growing bacterial cellulose in situ in wood fiber |
| CN114703185A (en) * | 2022-03-16 | 2022-07-05 | 天津科技大学 | Identification of the promoter function of the sequence derived from Acetobacter xylinum and its application in promoting bacterial cellulose synthesis |
| CN115287205A (en) * | 2022-05-17 | 2022-11-04 | 天津科技大学 | Schizosaccharomyces pombe with high acid resistance and construction method thereof |
| CN115927155A (en) * | 2022-12-19 | 2023-04-07 | 天津科技大学 | A screening method for strains resistant to lignocellulose-derived inhibitors |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101603059A (en) * | 2009-07-06 | 2009-12-16 | 江南大学 | A method for synchronous saccharification and fermentation of straw raw materials to produce succinic acid |
| CN101985641A (en) * | 2010-12-09 | 2011-03-16 | 东华大学 | Method for preparing bacterial cellulose by using wheat straw |
-
2017
- 2017-07-18 CN CN201710595452.2A patent/CN107164428A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101603059A (en) * | 2009-07-06 | 2009-12-16 | 江南大学 | A method for synchronous saccharification and fermentation of straw raw materials to produce succinic acid |
| CN101985641A (en) * | 2010-12-09 | 2011-03-16 | 东华大学 | Method for preparing bacterial cellulose by using wheat straw |
Non-Patent Citations (1)
| Title |
|---|
| 胡秋龙等: "木质纤维素生物质预处理技术的研究进展", 《中国农学通报》 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108315371A (en) * | 2018-04-20 | 2018-07-24 | 北京理工大学珠海学院 | A kind of cultural method of bacteria cellulose |
| CN109022534A (en) * | 2018-08-01 | 2018-12-18 | 淮阴师范学院 | Utilize the method and corn stover degradation product fuel cell of corn stover production bacteria cellulose film |
| CN109022534B (en) * | 2018-08-01 | 2020-06-30 | 淮阴师范学院 | Method for producing bacterial cellulose membrane by using corn straws and corn straw degradation product fuel cell |
| CN109182187A (en) * | 2018-09-20 | 2019-01-11 | 天津科技大学 | A method of utilizing glucose mass transfer in agar measurement Bacterial Cellulose in Fermentation Condition |
| CN110218751A (en) * | 2019-06-03 | 2019-09-10 | 广西大学 | The method for preparing cellobiose and cellooligosaccharide using carbohydrate |
| CN113913971B (en) * | 2020-07-10 | 2024-03-12 | 南京理工大学 | Method for in-situ growth of bacterial cellulose in wood fiber |
| CN113913971A (en) * | 2020-07-10 | 2022-01-11 | 南京理工大学 | A kind of method for growing bacterial cellulose in situ in wood fiber |
| CN113234637A (en) * | 2021-06-16 | 2021-08-10 | 南开大学 | Fermentation medium for large-scale efficient production of bacterial cellulose and fermentation method thereof |
| CN114703185B (en) * | 2022-03-16 | 2023-10-27 | 天津科技大学 | Application of acetobacter xylinum-derived promoter in promotion of bacterial cellulose synthesis |
| CN114703185A (en) * | 2022-03-16 | 2022-07-05 | 天津科技大学 | Identification of the promoter function of the sequence derived from Acetobacter xylinum and its application in promoting bacterial cellulose synthesis |
| CN115287205A (en) * | 2022-05-17 | 2022-11-04 | 天津科技大学 | Schizosaccharomyces pombe with high acid resistance and construction method thereof |
| CN115287205B (en) * | 2022-05-17 | 2024-02-06 | 天津科技大学 | Schizosaccharomyces pombe with high acid resistance and construction method thereof |
| CN115927155A (en) * | 2022-12-19 | 2023-04-07 | 天津科技大学 | A screening method for strains resistant to lignocellulose-derived inhibitors |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107164428A (en) | A kind of method for preparing bacteria cellulose as carbon source with ligocellulose degradation's liquid | |
| Kuo et al. | Hydrolysis of orange peel with cellulase and pectinase to produce bacterial cellulose using Gluconacetobacter xylinus | |
| Lin et al. | Production of bacterial cellulose by Gluconacetobacter hansenii CGMCC 3917 using only waste beer yeast as nutrient source | |
| CN103211291B (en) | Preparation method of reconstituted tobacco | |
| CN104945535B (en) | Method for production of sodium alginate and co-production of ethanol and seaweed organic fertilizer | |
| CN105255953B (en) | Physical-chemical-Biological Pretreatment corn stover method | |
| CN110172483A (en) | A method of ethyl alcohol is prepared using furfural dregs and cake of camellia oleifera seeds as raw material | |
| CN102660885A (en) | Method for preparing high-performance long fiber by utilizing lignocelluloses biological method | |
| CN101564680A (en) | Method for preparing compound biological adsorbent by solid-state fermentation of herb residue | |
| CN107828832A (en) | A kind of method for improving crudefiber crop stalk enzymatic saccharification efficiency | |
| CN114561327A (en) | Cellulose degradation composite microbial inoculum and preparation method and application thereof | |
| CN103352016A (en) | Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha | |
| CN109055457A (en) | A method of bacteria cellulose is produced using grape skin | |
| CN107177646B (en) | Method for strengthening acid pretreatment of waste biomass by using lignin-degrading bacteria | |
| CN108424941A (en) | A method of preparing bacteria cellulose film | |
| CN112401054A (en) | Continuous treatment method for lignocellulose raw material | |
| CN109486863B (en) | A method for degrading wood fibers in crop straw by using microbial inoculants | |
| CN107058427A (en) | A kind of method of utilization lignin-degrading bacteria reinforcing abandoned biomass ammonia process pretreatment | |
| CN108676194B (en) | A kind of composite polysaccharide hydrogel and its in situ biosynthesis method | |
| Beliah et al. | Utilization of marine algae as a carbon source for bacterial cellulose production by Gluconacetobacter xylinus | |
| CN102286572A (en) | A method for preparing fermentable sugar liquid using straw as raw material | |
| CN104946702A (en) | Method for pretreating lignocellulose raw material by combining ferric chloride and white rot fungi | |
| CN105002231A (en) | Method for preparing bacterial cellulose by biotransforming mulberry leaves | |
| CN111187627B (en) | A kind of low-cost, environment-friendly sand-fixing agent and preparation method thereof | |
| CN107287251A (en) | A kind of abandoned biomass method of resource |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170915 |
|
| WD01 | Invention patent application deemed withdrawn after publication |