111 results about "Clostridium acetobutylicum" patented technology

The invention provides a Clostridium acetobutylicum and applications thereof. The accession number of the Clostridium acetobutylicum provided by the invention is CGMCC No. 5234. The Clostridium acetobutylicum can be used to simultaneously produce acetone, butanol, ethanol and 3-hydroxybutanone through fermentation, so that economic benefits of butanol fermentation are increased. Coupled regeneration of NAD+ can be achieved by adding metabolism or growth regulators, which increases product yields and, at the same time, can flexibly adjust the yields of coproduction products to adapt to market demand.

The invention relates to the technical field of bio-fermentation, in particular to a clostridium acetobutylicum which can quickly use amylofermentation to produce biological butanol, and a screening method and application thereof; the invention discloses a strain of the clostridium acetobutylicum, which belongs to the mutant strain of the clostridium acetobutylicum CICC 8011 with the preservation number of CTCC M207199. The invention obtains the strain which can quickly use amylofermentation to produce biological butanol by NTG mutagenesis and double flat plate screening of starch transparent ring and analogs of the butanol, has high butanol rate, solves the problem that in the traditional technique, the waste mash COD is high by adopting the corn meal as materials and is more friendly to environment.

The invention provides a recombinant plasmid, a preparation method thereof, an application, two knockout recombinant plasmid vectors pSY6-buk and pSY6-solR which are prepared by taking the recombinant plasmid as a starting plasmid, a method for preparing the recombiant plasmid vectors pSY6-buk and pSY6-solR, and an application. The recombinant plasmid can be taken as a knonckout starting plasmid of an acetone and butanol, can perform the gene knockout efficiently and rapidly, and can be evolved into a novel genetic manipulation system in order to improve the imperfect condition of the prior manipulation system in the current research on the clostridium acetobutylicum, thereby providing convenience for the research on the metabolic engineering of the clostridium acetobutylicum. The recombinant plasmid vectors pSY6-buk and pSY6-solR can be directly applied to establishing knockout strains of the clostridium acetobutylicum buk and solR genes.

The invention discloses a genetically engineered bacterium for producing acetoin. The genetically engineered bacterium is clostridium into which a coding gene of acetolactate decarboxylic reaction enzyme is imported. The invention also discloses a construction method and an application of the genetically engineered bacterium. By adopting the method, the average yield of acetoin produced from clostridium acetobutylicum is about 5g/L.

The invention relates to a bacillus series straw fast rotting agent and a using method thereof. The fast rotting agent comprises bacillus licheniformis, bacillus circulans, bacillus subtilis, clostridium pasteurianum, bacillus megatherium, jelly-like bacillus, clostridium acetobutylicum, bacteria macerans, bacillus polymyxa and nutrient solution and is used for treating straws to make biological organic fertilizer. The using method comprises the following steps: stacking the straws into a walkway with the width of 40 cm and the height of 30 to 50 cm; uniformly spraying 1.25 kilograms of the fast rotting agent into per ton of straws; controlling the water content of the straws to be 40 to 60 wt%; and rotting at normal temperature for 1 to 2 weeks, wherein when the straws change into brown or black brown, the wet straws are flexible and elastic when being held by hands and the dry straws are crisp and easy to break, a straw decomposition agent product with the nutrient substance content up to 59 weight percent is obtained and can direct serve as fertilizer. By adoption of the fast rotting agent, the straw rotting time is reduced from 15-20 days at present to 7-10 days.

A genome-scale metabolic network reconstruction for Clostridium acetobutylicum (ATCC 824) was created using a new semi-automated reverse engineering algorithm. This invention includes algorithms and software that can reconstruct genome-scale metabolic networks for cell-types available through the Kyoto Encyclopedia of Genes and Genomes. This method can also be used to complete partial metabolic networks and cell signaling networks where adequate starting information base is available. The software may use a semi-automated approach which uses a priori knowledge of the cell-type from the user. Upon completion, the program output is a genome-scale stoichiometric matrix capable of cell growth in silico. The invention also includes methods for developing flux constraints and reducing the number of possible solutions to an under-determined system by applying specific proton flux states and identifying numerically-determined sub-systems. Although the model-building and analysis tools described in this invention were initially applied to C. acetobutylicum, the novel algorithms and software can be applied universally.

The invention discloses a method for ex-situ preparation of methane and combined production of humic acid, and a composite microbial agent used in the same, belonging to the field of recycling of brown coal. The composite microbial agent is composed of fermentation broth of four bacteria, i.e., Escherichia coli, Pseudomonas putida, Penicillium decumbens and Clostridium acetobutylicum. The composite microbial agent has strong degradation capability on a variety of celluloses, hydrocarbons, asphaltene and a variety of carbonaceous macro-molecules. As coal is treated by the composite microbial agent, anaerobic digestion time can be shortened, output of methane gas is increased and extraction efficiency of humic acid is improved; moreover, the synergistic effect of the above-mentioned four microbial strains is superior to the effect of any one microbial strain or any two microbial strains.

The invention discloses a method for regulating and controlling synthesis of an extracellular polymeric substance of clostridium acetobutylicum. The method is characterized in that an intron sequence is interpolated into a gene Spo0A of the clostridium acetobutylicum by using a type II intron gene knockout technology. The invention provides a simple and efficient method for regulating and controlling EPS (Extracellular Polymeric Substance) synthesis; a recombinant strain obtained based on the method is relatively good in regulating and controlling effect on the EPS; in case of taking glucose as a carbon source, when cultivated in an anaerobic bottle with the volume of 100 ml by using a slide biomembrane cultivation method, a clostisdium acetobutylicum biomembrane is 5.81 mum in thickness, and the thickness of a strain biomembrane cultivated under the same condition is 24.6 mum.

The invention discloses a microbial agent for sewage treatment. The microbial agent comprises, by weight, 8-10 parts of clostridium acetobutylicum, 12-16 parts of bacillus licheniformis, 16-20 parts of bacillus subtilis, 5-7 parts of lactobacillus acidophilus, 5-7 parts of methanosarcina, 3-5 parts of methanosaeta, 6-8 parts of rhodopseudomonas palustris, 4-6 parts of cellulomonas flavigena, 8-10parts of streptococcus thermophilus, 8-10 parts of alcaligenes faecalis, 3-5 parts of bacillus amyloliquefaciens, 2-4 parts of bacillus nitrosomonas, 1-3 parts of issatchenkia orientalis and 3-5 partsof streptomyces bullis. Besides, the invention further discloses an attachment carrier of the microbial agent and a preparation method of the microbial agent. The microbial agent can be effectively attached to the carrier and accordingly has better activity. The microbial agent is reasonable in strain preparation and can be effectively attached to the attachment carrier to treat sewage, and the method for sewage treatment is simple, good in treatment effect and worthy of popularization.

The invention relates to a Clostridium acetobutylicum strain and application thereof, belonging to the technical field of biofermentation. The classification designation of the Clostridium acetobutylicum strain provided by the invention is Clostridium acetobutylicum BD518, and the preservation registration number is CCTCC NO:M2010308. The high-concentration butanol-tolerant strain BD518 is obtained by continuous culture, domestication and screening, and the butanol tolerance is up to 18g/L and is increased by 38.5% in comparison with the original strain (13g/L). When glucose is used as a carbon source, the total solvent and butanol yields in a 5L fermentation tank are respectively up to 21.1g/L and 13.2g/L and are respectively increased by 16.6% and 15.8% in comparison with the original strain. The Clostridium acetobutylicum strain has high butanol tolerance, high solvent yield and good repeatability, thereby having significant social meanings and economic values.

The invention relates to a preparation method of peanut soy, belonging to the technical field of preparation methods of soy. The peanut soy provided by the invention adopts groundnut kernels as a raw material, bacillus licheniformis, brevibacterium flavum, clostridium acetobutylicum and rhamnose lactobacilli are taken as composite fermentative bacteria of a peanut fungi solution; the special proportional aspergillus oryzae, aspergillus niger and monascus are utilized as fermenting aspergillus of a peanut yeast, and a preparation process of natto soy is adopted. The preparation method provided by the invention has the advantages that the fermentation period is shortened, natto kinase is generated after fermenting of groundnut kernels, the kinase can restrain the browning of the soy in preservation, a maillard reaction is reduced, the nutritive values of the groundnut kernels are high, the flavor is unique, and the color, the aroma and the taste of the prepared soy are good; the content amino acid nitrogen is improved by 43.9%, the full nitrogen content is improved by 17.5%, a soluble salt-free solid is improved by 18.8%, and the preparation method is especially used for preparing ideal soy of pickles.

The invention discloses a genetically engineered bacterium for co-generating butanol and 2,3-butanediol. The genetically engineered bacterium is a butanol generating clostridium to which an acetoin reductase gene is introduced or a butanol generating clostridium to which an acetolactic acid decarboxylase gene and the acetoin reductase gene are introduced. The nucleotide sequence of the acetolactic acid decarboxylase gene is any one of nucleotide sequences shown in SEQ ID NO:1-7, and the nucleotide sequence of the acetoin reductase gene is any one of nucleotide sequences shown in SEQ ID NO:8-11. An acetolactic acid decarboxylic reaction is enhanced in a clostridium acetobutylicum, the yield of 3-hydroxy butanone is greatly increased to about 6.4 g/L from initial about 1.8 g/L, and then acetoin is converted into 2,3-butanediol reaching up to 8.05 g/L under the effect of acetoin reductase; acetone is reduced to 2.2 g/L from about 4.5 g/L, and economic benefits of fermentation of the clostridium acetobutylicum are remarkably increased.

The invention provides a method for extracting and isolating clostridium acetobutylicum exopolysaccharide, comprising the following steps: (1), inoculating clostridium acetobutylicum seed liquid to a fermentation tank, adding mobilizing material, culturing until the surface of the mobilizing material is fully covered by clostridium acetobutylicum biofilm; (2), dissolving clostridium acetobutylicum biofilm with alkaline solution, centrifuging to remove bacterial precipitate, adding an organic solvent into supernate so that the organic solvent is 30-90% by weight, the organic solvent being ethanol, acetone or butanol, centrifuging, and collecting precipitate; adding an acid to adjust pH to 3.5-10.0, and collecting precipitate, the precipitates collected twice being the exopolysaccharide. The method provided herein has the advantages of low cost and good operation simplicity.

The invention discloses clostridium acetobutylicum for producing butanol by utilizing manihot as raw materials and application thereof. The clostridium acetobutylicum is preserved in China Center for Type CultureCollection with the number of CCTCC M 2011280. The clostridium acetobutylicum is capable of fermenting to produce the butanol, and particularly, the fermentation volume of the butanol in a fermentation medium taking the manihot as the raw materials can reach 13.07 g/L.

The invention relates to a method for preparing n-butyl alcohol, in particular to a method for preparing n-butyl alcohol in a fermentation mode by hydrolyzing lignocellulose through mixed cellulose crude enzymes. According to the method, the problems that in the existing process that the n-butyl alcohol is produced by biological fermentation, a substrate is high in cost, the production cost is increased due to the use of commercial enzymes in the hydrolytic process of the lignocellulose, and the enzyme activity is reduced due to an unbalanced proportion of a cellulose system of the single kind of culture are solved. The method comprises the steps that the preprocessed lignocellulose is applied to the mixed cellulase crude enzymes of trichoderma viride and aspergillus niger and hydrolysis is conducted according to a biological enzyme method; after a nitrogen source, inorganic salt and vitamins are added to obtained liquid glucose, nitrogen is added to the liquid for deoxygenization; a clostridium acetobutylicum seed solution is inoculated for anaerobic fermentation, and therefore the n-butyl alcohol is generated. According to the method, the activity of biological enzymes is improved in the cellulose enzymolysis process, cost of the substrate and the enzymatic hydrolysis in the process that the n-butyl alcohol is produced by biological fermentation is reduced, the problems of device losses and environment pollution are solved, and the saccharity hydrolysis efficiency of the mixed crude enzymes reaches 75%.

The invention discloses recombinant corynebacterium glutamicum in high L-lysine yield and a construction method thereof, and belongs to the technical field of genetic engineering. According to the recombinant corynebacterium glutamicum disclosed by the invention, a phosphoglyceraldehyde dehydrogenase encoding gene gapC derived from Clostridium acetobutylicum ATCC824 is heterogeneously expressed bytaking corynebacterium glutamicum CICC 23604 as an original strain and adopting CRISPR-Cas9 gene editing technology, and supply of intracellular NADPH (Nicotinamide Adenine Dinucleotide Phosphate) isincreased; a homoserine kinase encoding gene thrB is knocked out, accumulation of a byproduct-threonine during a fermentation process is reduced, corynebacterium glutamicum genetically engineered bacteria with the accumulation of L-lysine can be obtained, the yield is up to 42g/L, and foundation is laid for producing the L-lysine through further metabolism engineering reform of the corynebacterium glutamicum.

The present invention is related to a population of Clostridium acetobutylicum useful for the production of 1,3-propanediol (PDO), wherein said population comprises at least one strain of a Clostridium acetobutylicum sp. comprising mutations selected among the mutations identified in table 1, wherein relative percentages of said mutations are selected among specific genes.

A cell immobilization method of producing butanol clostridium adopts the butanol clostridium liquor at exponential growth phase to produce immobilized cells through the adsorption of straw. The method includes main steps as follows: A) crushing the corn stalk into small pieces, bathing in the dilute hydrochloric acid or dilute sulfuric acid at 80-100 DEG C for boiling 10-60 minutes, cleansing, and drying to a constant weight; B) immerging the corn stalk processed by the step A into a butanol clostridium cell culture fluid at exponential growth phase, static adsorption at 30 -40 DEG C for 12-48 hours. The inventive immobilization method has features of low cost, effective cell-rich effect, high activity of immobilized cells, and high production efficiency during continuous fermentation.

The invention relates to a Clostridium acetobutylicum strain capable of quickly generating butanol by fermenting xylose and starch, and a screening method and application thereof. The invention discloses Clostridium acetobutylicum, which is classified and named Clostridium acetobutylicumXY16 with the preservation and registration number of CCTCC NO:M2010011. The strain obtained by ion beam mutation and screening of a xylose plate, a 2-deoxy-D-glucose plate, a bromocresol green plate and a resazurin plate can quickly and efficiently generate the butanol by fermenting xylose and starch, has the advantages of high yield of total solvent, high butanol ratio and good repeatability, and is a good strain suitable for industrial production.