33 results about "Glycerol fermentation" patented technology

The present invention is in the field of producing organic acids and other useful chemicals via biological fermentation using glycerol as a source of carbon. Novel microorganisms and fermentation processes are described that are capable of converting glycerol to useful organic acids in high yield and high purity.

A method for preparing phytase using glycerol as a carbon source, adopting the fermentation production process of phytase-producing genetically engineered bacteria using Pichia pastoris as a carrier, characterized in that glycerol is used as a carbon source, and an appropriate amount of bacterial strains for production is used , cultured in a shake flask in a seed medium, and then through the first-level seed culture, second-level seed culture, and fermentation culture processes, the only carbon source in the growth stage of the bacteria before the seed culture and fermentation enzyme production is glycerol; during fermentation, After the glycerol in the basic fermentation medium is completely consumed, start to feed 50% glycerol solution into the medium. After the glycerol is exhausted, start to feed methanol to induce phytase production. Methanol also acts as a carbon source in the production stage. The positive effect is: Glycerin as a carbon source increases the final phytase production by 20-30%, increases the enzyme activity by 20-30%, the color of the fermentation liquid is light and does not require decolorization treatment, and reduces production costs. This method reduces costs compared with traditional processes About 1 / 3.

The invention relates to a method for producing sophorose ester through fermenting waste molasses and waste glycerin, which comprises the following implementation steps that: (1) spore suspension preparation: torulopsis utilis TJZKBA 10326 is inoculated on the inclined surface of a culture medium and is cultured for 2 to 4 days at the constant temperature being 30 DEG C, and normal saline with the mass concentration being 0.75 percent is added for washing mycelia, and the bacterial suspension is obtained; (2) the bacterial suspension is used for being inoculated into the sterilized liquid seed culture medium, the spore suspension is inoculated into the cooled liquid seed culture medium according to the inoculation quantity with the volume percentage being 2 to 5, and liquid seeds are obtained; (3) the liquid seed culture medium is used for being inoculated into a sterilized 5L fermentation tank, then, the liquid culture medium is inoculated into the cooled fermentation tank culture medium according to the inoculation quantity with the volume percentage being 2 to 5, the fermentation temperature is 30 to 34 DEG C, the dissolved oxygen is controlled to be 30 to 40 percent, and the fermentation time is 48 to 72h; and (4) after the fermentation is completed, the mixed organic solvent of organic solvent ethyl acetate / isopropanol with the same volume as the fermentation liquid is used for extracting the sophorose ester. The method has the advantages that the process is simple, the cost is low, and the effect is obvious.

The invention discloses a method for producing 1,3-propanediol and 2,3-butanediol from raw starch materials, including the following steps: 1) Candida krusei or Hansenula Arabitolgens Fang are inoculated into a fermentation medium with the saccharifying liquid of the raw starches as a carbon source; the yeast cells are cultured on an aerobic condition until glucose-consuming-rate is significantly reduced, and then fermented anaerobically to a glucose concentration from 5 to 10 g / L; the fermentation broth is collected and filtered to remove the yeast cells in the broth, and the resultant filtrate is glycerin fermentation broth; 2) Klebsiella, Clostridium butyricum, or Clostridium pasteurianum are inoculated into a fermentation medium in which the glycerin fermentation broth obtained from step 1) serves as a carbon source; the bacteria are fermented anaerobically for 30-32 hours, and then fermented aerobically when the production rate of 1,3-propanediol decreased obviously, and the fermentation was stopped when the concentration of glycerin is reduced to a level below 10 g / L, and finally 1,3-propanediol and 2,3-butanediol are obtained. The method of the present invention can effectively reduce production cost and increase productivity.

The invention relates to the synthesis of a 1,3-dioxyacetone. A method for synthesizing 1,3-dioxyacetone by glycerol fermentation comprises the following steps: performing the gene recombination and DNA rearrangement technology on bacillus licheniformis B-05571 separated and screened from acid soil, integrating a dehydrogenase gene into an Escherichia coli JM109, constructing industrial engineering bacteria of 1,3-dioxyacetone synthesized by glycerol dehydrogenization, immobilizing by using a supermolecule self-assembly template, then fermenting in a culture medium containing glycerol to synthesize 1,3-dioxyacetone. The product 1,3-dioxyacetone can be obtained by the steps of filtering fermentation liquor, extracting with an organic solvent and recrystallizing with a combined solvent. The synthesis process route is simple and convenient, the product is high in quality and low in cost, and the total mass yield is higher than 75%.

The invention discloses a genetic engineering strain capable of coproducing isoprene and 1,3-propylene glycol and an establishment method and application thereof, and belongs to the technical field of genetic engineering. In the genetic engineering strain, a glycerol dehydratase gene and a glycerol dehydratase gene reactivating enzyme gene are integrated, a glycerol kinase gene is knocked out, and HMG-CoA reductase, HMG-CoA synthetase, mevalonate kinase, phosphomevalonate kinase, pyrophosphomevalonate decarboxylase, IPP isomerase, isoprene synthetase, aldehyde reductase and transhydrogenase are overexpressed. According to the invention, through genetic engineering, a metabolic way capable of coproducing the isoprene and the 1,3-propylene glycol is successfully established in escherichia coli; by the establishment method, redox cofactors in cells of the genetic engineering strain are metabolized in a balanced way, and the yield in coproducing the isoprene and the 1,3-propylene glycol through glucose and glycerol fermentation is increased.

The invention belongs to the technical field of bioengineering, and particularly relates to a clostridium butyricum strain with high yield of 1,3-propanediol and a sequential inoculation fermentationprocess thereof. The strain is named as clostridium butyricum DL07 and a preservation number is CGMCCCNO. 17934. The strain of the present invention can produce 104.78 g / L and 94.23 g / L of 1,3-propanediol by fermentation with pure glycerin and crude glycerin respectively, and the currently reported natural microorganisms uses the pure glycerin and crude glycerin to ferment and produce the highestyield of 1,3-propanediol. The invention also provides a sequential inoculation fermentation process and an automatic fed-substrate fermentation control method, which saves seed cultivation time and realizes multi-stage continuous fermentation, and provides a new fermentation control mode with high degree of automation and excellent economy for the production of 1,3-propanediol by microbial fermentation, and the process has the good industrial application prospect.

The invention discloses clostridium butyricum and an application thereof in immobilized fermentation production of 1, 3-propylene glycol, the clostridium butyricum is clostridium butyricum SCUT343-4 and is preserved in Guangdong province microbial strain preservation center on October 12, 2019, and the preservation number is GDMCC NO: 60805. The strain can be used for directly converting pure glycerol or crude glycerol into 1, 3-propylene glycol, has higher substrate conversion rate and substrate tolerance, has few types and contents of byproducts, and is beneficial to separation and purification of subsequent products; the strain has the most outstanding tolerance to a substrate crude glycerol, and the conversion rate and the production efficiency are not reduced during crude glycerol fermentation, so that the processing and utilization of the crude glycerol are facilitated, and the production cost of the 1, 3-propylene glycol is remarkably reduced. The strain is applied to immobilized fermentation production of 1, 3-propylene glycol, the production efficiency can be remarkably improved and reaches 4.20 g / L.h.

The invention discloses a production technology of 1,3-propylene glycol (PDO) by using a biological fermentation method, belonging to the technical field of biochemical engineering. The production technology comprises the following steps of: preparing 2'-dehydroxylase gene (sisII) from 3',4'-double dehydroxylase gene of sisI in micromonospora inyoensis through genetic recombination, DNA (deoxyribonucleic acid) rearrangement and the like; cloning to obtain sisII; constructing the sisII to a pET30a expression vector, and performing heterologous expression in E.coli BL21 to construct engineering bacteria BL21-sis for synthesizing 1,3-propylene glycol through glycerol dehydration; performing immobilization by a supramolecular self-assembly template method; fermenting to synthesize 1,3-propylene glycol in a glycerol-containing culture medium; and obtaining the product 1,3-propylene glycol through fermentation liquid filtration, organic solvent extraction and the like. The method for preparing 1,3-propylene glycol has the advantages of short production period, simple equipment, environmental friendliness, easiness in operation and control, cheap raw materials and the like, facilitates industrial production and has great practical value.

The invention provides a method for producing ethanol by adopting paenibacillus macerans and monasucs by means of glycerol mixed culture fermentation. According to the invention, the paenibacillus macerans and monasucs are taken as original strains and glycerol as raw materials, researches are carried out on initial glycerol concentration, glycerol concentration in feeding supplement, feeding supplement time interval, fermentation time and slurry quantity that influence the ethanol yield.

Glycerol fermentation by candida glycerinogenes NAD+ depends on glycerine-3-phosphate dehydrogenase gene and clone thereof and pertains to the microorganism molecular biology field. The invention relates to a new gene order SEQID NO.1. a degenerate primer PCR method and a reverse PCR method are adopted to clone rate-limiting enzyme NAD+ depending glycerine-3-phosphate dehydrogenase gene and lateral regulation sequence thereof from genome DNA of glycerine-producing candida mycoderma WL2002-5-59 which is produced by glycerine. The total length of the gene is 4900bp and a start codon ATG is at 2082bp while a stop codon TAA is at 3246bp. The sequence has no intron but has an 1167bp complete open reading frame which encodes 388 amino acids. The gene has homology as high as 60 percent with saccharomyces cerevisia GPD1 gene and 70 percent with Angus pichia yeast GPD gene. The clone of the gene expands gene resource of microorganism anti-osmotic pressure stress research and lays a foundation for research on molecular mechanism of glycerine-producing candida mycoderma producing glycerine with high yield.

The invention provides a method for producing 1,3-propanediol by fermentation of glycerol in a microbial co-cultivation system, by successfully constructing recombinant Escherichia coli Rosseta-dhaB1dhaB2 and Rosseta-dhaT, co-cultivating the above-mentioned recombinant Escherichia coli for seed culture and fermentation culture , wherein the recombinant Escherichia coli Rosseta-dhaB1dhaB2 realizes the utilization of glycerol to produce the intermediate metabolite 3-hydroxypropionaldehyde (3-HPA), and the recombinant Escherichia coli Rosseta-dhaT catalyzes 3-HPA to produce 1,3-propanediol. During the fermentation process, press Inoculate in the fermentation medium at a ratio of 1:1.5, add 60g / L substrate glycerol and 10g / L cosubstrate glucose. The method of the present invention improves the conversion rate of glycerol and the output of 1,3-propanediol, and the present invention adopts Clostridium butyricum containing independent coenzyme B 12 Glycerol dehydratase, which reduces the production cost.

The invention relates to the technical field of engineered strain construction, in particular to a construction method of an engineered strain for producing phenol through glycerol fermentation. The second base sequence and the third base sequence in a sequence list are inserted in a pTrc99a carrier to obtain recombinant plasmid; the recombinant plasmid is electrically transformed into an ATC31884 strain, the engineered strain for producing the phenol through glycerol fermentation is obtained. The constructed engineered strain efficiently presents tyrosine phenol lyase and phenylalanine4-monooxygenase, and content of the obtained escherichia coli engineered strain for producing the phenol through glycerol is 21.30-57.66 mg / L, so that the phenol is produced through glycerol fermentation through the biological method, production cost of the phenol is reduced, and economic benefits are improved.

The invention provides a method for producing 1,3-butanediol through glycerol fermentation via a microbial co-culture system. The method comprises the following steps: successfully constructing recombinant escherichia coli Rosseta-dhaB1dhaB2 and Rosseta-dhaT, co-culturing the recombinant escherichia coli, and carrying out seed culture and fermentation culture, wherein the recombinant escherichia coli Rosseta-dhaB1dhaB2 is used for realizing the production of the intermediate metabolite 3-hydroxypropionaldehyde (3-HPA) from glycerol, the recombinant escherichia coli Rosseta-dhaT catalyzes 3-HPAto produce 1,3-propanediol, and in the process of fermentation, a fermentation medium is inoculated with the recombinant escherichia coli in a ratio of 1: 1.5, and a substrate glycerinum with a concentration of 60 g / L of and an auxiliary substrate glucose with a concentration of 10 g / L of are added. According to the method disclosed by the invention, the conversion rate of glycerol and the yieldof 1,3-propylene glycol are increased; and the adopted clostridium butyricum contains glycerol dehydratase which does not depend on coenzyme B12, so the production cost is reduced.

The invention relates to a method for producing 1,3-propylene glycol by glycerol fermentation. The method comprises the following steps: firstly, culturing fermentation strains under a microaerobic condition to obtain a fermentation seed liquid; then, culturing microalgae under an autotrophic condition to obtain a microalgae seed liquid; then, inoculating a fermentation culture medium with the fermentation seed liquid to carry out microaerobic fermentation, adding the microalgae seed liquid when the acetic acid concentration is more than 3g / L, and performing culturing until fermentation is finished; and finally, adjusting the pH value to be alkaline, and performing standing for layering to obtain fermentation clear liquid containing 1,3-propylene glycol. According to the method provided bythe invention, through combination of microalgae cell metabolism and a fermentation system, oxidative phosphorylation of the substrate level in glycerol metabolism is strengthened, and the fermentation level is improved.

The invention discloses pseudomonas aeruginosa as well as a construction method and application thereof. The invention discloses a pseudomonas aeruginosa recombinant bacterium which is used for overexpressing NAD (Nicotinamide Adenine Dinucleotide) + synthetase. The invention provides a new development strategy for producing rhamnolipid by using microorganisms through glycerol fermentation.

The invention discloses a bacterial strain that strengthens glycerol metabolism, and the bacterial strain is classified and named as Actinobacillus succinate ( Actinobacillus succinogenes ) JF1315, which was deposited in China Center for Type Culture Collection on March 31, 2016, and its deposit number is: CCTCC NO: M 2016160. The present invention also relates to the application of the Actinobacillus succinogenes JF1315 in fermentative production of organic acids. Under the anaerobic condition, compared with the control strain, the strain can grow and metabolize at a lower pH condition, and can efficiently utilize glycerol to ferment and synthesize reduced organic acids. Under the condition of pH 4.8-6.8, the strain can grow normally, and metabolize glucose to synthesize organic acids such as succinic acid; in addition, under this condition, the strain can efficiently use glycerol for anaerobic fermentation, synthesize and accumulate organic acids. acid.

The invention relates to the technical field of engineered strain construction, in particular to a construction method of an engineered strain for producing phenol through glycerol fermentation. The second base sequence and the third base sequence in a sequence list are inserted in a pTrc99a carrier to obtain recombinant plasmid; the recombinant plasmid is electrically transformed into an ATC31884 strain, the engineered strain for producing the phenol through glycerol fermentation is obtained. The constructed engineered strain efficiently presents tyrosine phenol lyase and phenylalanine4-monooxygenase, and content of the obtained escherichia coli engineered strain for producing the phenol through glycerol is 21.30-57.66 mg / L, so that the phenol is produced through glycerol fermentation through the biological method, production cost of the phenol is reduced, and economic benefits are improved.

The invention relates to a method for producing 1,3-propanediol by fermentation of glycerin. Firstly, the fermentation strain is cultivated under microaerobic conditions to obtain a fermented seed liquid; then microalgae are cultured under autotrophic conditions to obtain a microalgae seed liquid Then, insert the fermented seed liquid in the fermentation medium to carry out micro-aerobic fermentation, add the microalgae seed liquid when the acetic acid concentration is greater than 3g / L, and cultivate until the end of fermentation; finally adjust the pH to alkaline, and let it stand for stratification A fermentation broth containing 1,3-propanediol is obtained. The method of the invention utilizes the combination of microalgae cell metabolism and fermentation system to strengthen the oxidative phosphorylation at the substrate level in glycerin metabolism and improve the fermentation level.

The invention provides a method for mixed culture of R.salina; the method includes the steps: culturing R.salina with a seed culture medium; carrying out domestication culture by using glycerol; inoculating a glycerol fermentation culture medium with the R.salina after domestication; and culturing the R.salina by using light with different wavelengths and intensities. The thallus substrate utilization rate, growth rate, living weight, and phycoerythrin, total protein, polyunsaturated fatty acid and total lipid contents during R.salina fermentation are improved, the problems of harvest of the R.salina are reduced, the biomass concentration and the pre-processing cost of target product extraction are reduced.

The invention discloses a production method and application of mannanase. The method for producing mannanase provided by the present invention comprises: introducing the mannanase gene derived from Cladosporium camphoropsis S168 into recipient yeast to obtain recombinant yeast; performing basic fermentation culture, glycerol In the fed-feed fermentation stage and the methanol-feed-induced expression stage, the mannanase was obtained from the fermentation product. The mannanase expression level of the engineering bacteria of the present invention reaches 42200U / mL through high-density fermentation, and the optimal pH of the obtained enzyme is 7.5, and the optimal temperature is 75°C. It has acid and alkali resistance, good thermal stability, and excellent hydrolysis properties. Features, in addition, the present invention broadens the scope of application of mannanase, and the partially hydrolyzed konjac gum prepared by using this enzyme has great application value in industries such as food and feed, enriches the types of prebiotic products, and promotes the development of prebiotic products in my country. The development of yuan industry has great social and economic benefits.

The invention discloses a bio-fermentation method for preparing 1,3-propanediol. The method comprises the following steps: performing aerobic and anaerobic fermentation on crude starch as a raw material in a No. 1 fermentation tank by adopting pressure-resistant yeast to obtain glycerol fermentation broth, filtering the fermentation broth, recovering thalli for next fermentation, transferring a filtrate into a No. 2 fermentation tank accommodating Klebsiella pneumoniae for anaerobic fermentation, performing flocculating, filtering, evaporating and concentrating, reduced pressure distillation and decolorization on the fermentation broth to obtain the finished product 1,3-propanediol. With application of the method, the technical level of bio-fermentation in China is greatly improved, raw materials can be used to the greatest extent, the production cost is effectively reduced, productivity of 1,3-propanediol is improved, and the method is more suitable for industrial production.

The invention discloses the construction and application of an Escherichia coli engineering bacterium producing n-butyric acid, and belongs to the technical field of fermentation engineering. The present invention takes Escherichia coli ATCC 8739 as the starting strain, utilizes metabolic engineering means, constructs butyric acid-producing Escherichia coli genetically engineered bacteria by constructing a butyric acid synthesis path and cutting off redundant metabolic branches. After 72 hours of fermentation, the production of butyric acid reached 15g / L. The fermentation process is an aerobic fermentation, the bacteria grow fast, the fermentation period is short, and the acid production rate is high. At present, there is no report of directly using glycerol to ferment and produce butyric acid. The method adopts a simple fermentation process, is easy to control, has low production cost, and is beneficial to popularization and application of industrial production.