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9 results about "Clostridium thermocellum" patented technology
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Clostridium thermocellum is an anaerobic, thermophilic bacterium. C. thermocellum has garnered research interest due to its cellulolytic and ethanologenic abilities, being capable of directly converting a cellulosic substrate into ethanol by consolidated bioprocessing. This makes it useful in converting biomass into a usable energy source. The degradation of the cellulose is carried out in the bacterium by a large extracellular cellulase system called a cellulosome, which contains nearly 20 catalytic subunits. The cellulase system of the bacterium significantly differs from fungal cellulases due to its high activity on crystalline cellulose, being able to completely solubilize crystalline sources of cellulose, such as cotton. However, there are some shortfalls in applying the organism to practical applications due to it having low ethanol yield, at least partially due to branched fermentation pathways that produce acetate, formate, and lactate along with ethanol. There is also evidence of inhibition due to the presence of hydrogen and due to agitation. Some recent research has been directed to optimizing the ethanol-producing metabolic pathway in hopes of creating more efficient biomass conversion.
Clostridium thermocellum engineering strain with high nitrogen utilization rate and construction and application of clostridium thermocellum engineering strain
The invention provides a clostridium thermocellum engineering strain with a high nitrogen utilization rate. NitR regulatory protein is precisely overexpressed in the engineering strain; the precise overexpression is as follows: the expression level of the regulatory protein NitR is 1.2-2.0 times that of a wild type. A chassis strain of the strain is clostridium thermocellum GB2 with a glucosidase expression cassette on a genome, and the clostridium thermocellum GB2 is an engineering strain for expressing BGL in clostridium thermocellum DSM1313. The inventor accidentally finds that when the expression level of the regulatory protein NitR is controlled to be 1.2-2.0 times that of a wild type, the growth ability and biomass accumulation of the engineering strain under the nitrogen-deficient condition are remarkably improved, so that the utilization efficiency of a nitrogen source is increased, and an unexpected technical effect is achieved. The invention further provides a construction method of the clostridium thermocellum engineering strain and application of the clostridium thermocellum engineering strain in saccharification of lignocellulose raw materials. The engineering strain is particularly suitable for saccharification treatment of low-nitrogen-content lignocellulose raw materials (such as corn straw and corn cob residues), the saccharification efficiency is improved while the nitrogen source adding cost is remarkably reduced, and the engineering strain has important significance on industrial application.
Owner:QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
Clostridium thermocellum engineering strain for resisting phenolic hydroxyl inhibitors as well as construction and application of clostridium thermocellum engineering strain
The invention provides a clostridium thermocellum engineering strain for resisting phenolic hydroxyl inhibitors. Aldoketoreductase 1946 or aldoketoreductase 1992 is overexpressed in the clostridium thermocellum engineering strain at the same time; a chassis strain of the strain is clostridium thermocellum GB2 obtained by integrating a glucosidase expression cassette onto a DSM1313 strain genome. The invention provides two construction methods of a clostridium thermocellum engineering strain for resisting phenolic hydroxyl inhibitors, namely subculture: subculturing clostridium thermocellum in a vanillin-containing culture medium for 1-3 times to obtain a resistant strain with a delay phase of at least 3 hours; or directional transformation: constructing a recombinant strain through high-expression aldoketoreductase. The engineering strain relieves the inhibition of phenolic hydroxyl inhibitors generated in the lignocellulose pretreatment process on clostridium thermocellum, and has important practical application value. The invention further provides application of the clostridium thermocellum engineering strain in lignocellulose whole-cell saccharification, so that the saccharification efficiency is remarkably improved, and the technical effect is remarkable.
Owner:QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
A type of Co-added 2+ Methods to improve the yield of D-allulose produced by multi-enzyme catalysis
The application discloses a kind of added Co 2+ The application discloses a method for improving the yield of D-allulose produced by multi-enzyme catalysis, belonging to the field of biotechnology. The application provides a method for producing D-allulose by multi-enzyme catalysis, which comprises adding sucrose phosphorylase from Bifidobacterium adolescentis, fructokinase from Clostridium acetobutylicum, D-allulose-6-phosphate phosphatase from Clostridium thermocellum, and D-allulose-6-phosphate-3-epimerase from Pantoea sp to a reaction system containing sucrose and ATP, and then adding a metal ion solution to the reaction system to produce D-allulose. The application synthesizes an important low-calorie functional rare sugar D-allulose using inexpensive sucrose as a substrate, and experiments show that the addition of cobalt ions can increase the yield of D-allulose by five times.
Owner:JIANGNAN UNIV
A treatment method for low-concentration PCB copper-containing wastewater based on anaerobic conditions
The present invention relates to the technical field of sewage treatment, and specifically discloses a method for treating low-concentration PCB copper-containing wastewater under anaerobic conditions. The method includes the following steps: Step 1), inoculating Clostridium thermocellum, Thermoanaerobacter ethanolicus, and Rhodopseudomonas palustris into a nutrient solution and performing anaerobic culture to obtain a wastewater treatment bacterial solution; Step 2), injecting the copper-containing wastewater into a container, adding the wastewater treatment bacterial solution, and performing anaerobic treatment for 7-10 days to obtain pretreated wastewater; Step 3), filtering and sterilizing the pretreated wastewater to obtain pre-copper-removed wastewater. The present invention has the advantages of reducing the cost of the copper removal process for PCB wastewater and having relatively high economic value.
Owner:MEIZHOU HUAYU SEWAGE TREATMENT CO LTD +1
Extracellular cellulose response type post-transcriptional regulatory element suitable for clostridium thermocellum and application thereof
The invention innovatively provides a novel cellulose response type posttranscriptional regulatory element based on RNA conformation change. The regulation element comprises a core hairpin structural domain and a downstream terminator structural domain which are connected in series; the nucleotide sequence of the core hairpin structural domain is as shown in SEQ ID NO: 1, and the nucleotide sequence of the downstream terminator structural domain is as shown in SEQ ID NO: 2. The cellulose response type post-transcriptional regulation element disclosed by the invention has high-temperature applicability, can ensure the stability and responsiveness of a hairpin structure and a terminator at high temperature at the optimal growth temperature (55-60 DEG C) of clostridium thermocellum, and solves the core problem of scarcity of inducible tools in high-temperature chassis bacteria. On the basis, the invention further provides an engineering strain, the problem that the thermophilic bacterium clostridium thermocellum lacks a substrate induction type expression tool is solved, precise coupling of gene expression and a cellulose substrate is achieved, and the saccharification efficiency of lignocellulose and the metabolic engineering application potential can be remarkably improved.
Owner:QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
A method for hydrogen production by degrading undetoxified biomass and its application
The present invention relates to the technical field of biomass hydrogen production, and specifically discloses a method for degrading undetoxified biomass to produce hydrogen and its application. It includes the following operating steps: Step 1: Preparation of seed culture solution: Thermoanaerobacterium thermosaccharolyticum and Clostridium thermocellum are respectively inoculated into seed medium I and seed medium II, activated and expanded; Seed culture solution I and seed culture solution II are respectively obtained; Step 2: Preparation of fermentation medium: (1) Waste biomass is pretreated in a pretreatment solution, and solid-liquid separation is carried out to obtain pretreated biomass; (2) Using the pretreated biomass as a carbon source, a fermentation medium is prepared; Step 3: Under an inert gas, after sterilization, seed culture solution I and seed culture solution II are inoculated into the fermentation medium in a certain proportion, and dark fermentation is carried out at 50-60°C and a rotation speed of 100-200 rpm for 16-32 h to obtain hydrogen.
Owner:DONGGUAN UNIV OF TECH
A genetically engineered strain for high-yield d-allulose and a construction method and application thereof
This invention relates to the fields of synthetic biology and metabolic engineering, and particularly to a genetically engineered strain producing high levels of D-allulose, its construction method, and its applications. The strain uses *Escherichia coli* as the starting strain and constructs a phosphorylation / epimerization / dephosphorylation synthetic pathway by introducing *AlsE* from *E. coli* and *CtA6PP* from *Clostridium thermocellum*. Competitive metabolic genes *pgm*, *zwf*, *pfkA*, and *rpiB* are knocked out to enhance carbon flux directing. Furthermore, the expression of *AlsE* and *CtA6PP* is precisely regulated at the transcriptional and translational levels through promoter engineering and RBS engineering to obtain the optimal expression combination. The engineered strain achieves a D-allulose yield of 5.26 g / L in in vitro fermentation and 33.05 g / L in a 5L fed-batch fermenter, exhibiting advantages of high yield, stability, and ease of scale-up, making it suitable for industrial production of D-allulose.
Owner:HENAN ZHONGDA HENGYUAN BIOTECH CO LTD
Thermophilic L-arabinose inducible gene regulation system and application thereof
PendingCN120989081AMicroorganism based processesBacteria peptidesRegulation of gene expressionClostridium organisms
The invention provides a thermophilic L-arabinose inducible gene regulation and control system suitable for clostridium thermocellum as well as construction and application of the thermophilic L-arabinose inducible gene regulation and control system. The inducible gene regulation system comprises (a) an AraR inhibitory protein and a strong promoter for controlling the expression of the protein, (b) an inducible promoter regulated by the AraR inhibitory protein, and (c) a reporter gene operably connected to the inducible promoter. As the transformation of an expression promoter in the AraR inhibitory protein significantly improves the induction multiple and reduces leakage expression, the thermophilic inducible gene expression regulation system disclosed by the invention has excellent leakproofness, inductivity and dynamic range; the technical blank that no high-performance induction system suitable for high-temperature strains exists in the prior art is filled. In addition, an inducer adopted by the induction system is L-arabinose; the inducer simultaneously meets two conditions of low cost and stable inducible expression, and has important significance for practical application. The invention further provides application of the inducible gene expression regulation system in other strains, the strains are high-temperature strains or medium-temperature strains, and the application range of the inducible gene expression regulation system is further expanded.
Owner:QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
Method for synthesizing UDP-glucose through cooperation of UDP-glucose pyrophosphorylase and cellobiose phosphorylase and one-pot method
PendingCN121992050ASimple processImprove thermal stabilityTransferasesFermentationCellobiose phosphorylaseUDPglucose pyrophosphorylase
The invention discloses a method for synthesizing UDP-glucose through cooperation of UDP-glucose pyrophosphorylase and cellobiose phosphorylase by a one-pot method, and belongs to the technical field of enzyme engineering. The method comprises the following steps: selecting UDP-glucose pyrophosphorylase (UGPase) from mycelia thermophilus and cellobiose phosphorylase (CBP) from clostridium thermocellum, and carrying out catalytic reaction by taking cellobiose and UTP as raw materials, so as to obtain UDP-glucose. Experimental results show that heat-resistant UGPase is coupled with CBP, substrates cellobiose and UTP can be efficiently and highly selectively converted into UDP-glucose under mild conditions, and accumulation of an intermediate product glucose-1-phosphoric acid is extremely low. According to the invention, a synergistic reaction system with matched conditions is constructed, and a novel and efficient method for synthesizing UDP-glucose by a one-pot method is provided.
Owner:SHANDONG UNIV