Abstract: The present invention discloses a RamA transcription factor mutant for promoting the production of N-acetylglucosamine and use thereof. The mutant is obtained by mutating lysine at position 90 to asparagine and serine at position 92 to lysine in a parent having an amino acid sequence as shown in SEQ ID NO: 2. The present invention provides a genetically engineered strain that overexpresses the RamA transcription factor mutant and increases the production of N-acetylglucosamine. By overexpressing the transcription factor RamA that is involved in the regulation of carbon metabolism, the extracellular accumulation of N-acetylglucosamine is increased, with a maximum concentration reaching 31.5 g/L, which lays a foundation for further metabolic engineering of Corynebacterium glutamicum to produce glucosamine. The method for constructing recombinant Corynebacterium glutamicum of the invention is simple, and convenient in use, and thus has good application prospects.

Abstract: The present invention provides a recombinant Bacillus subtilis with improved 2?-fucosyllactose production, and a construction method thereof. In the present invention, a strain capable of efficiently synthesizing 2?-fucosyllactose is obtained by the fusion expression of the fucosyltransferase gene and the L-fucokinase/guanosine 5?-diphosphate-L-fucose pyrophosphorylase gene in Bacillus subtilis BSGL-FF, the fermentation supernatant of which comprises a cumulative amount of 2?-fucosyllactose as high as 1.62 g/L, which is 55% higher than the amount achieved with the control strain. The construction method of the recombinant Bacillus subtilis of the present invention is simple, and convenient to use, and thus has good application prospects.

Abstract: The present invention provides a recombinant Corynebacterium glutamicum producing N-acetylglucosamine and use thereof. The recombinant Corynebacterium glutamicum is obtained by overexpressing, in Corynebacterium glutamicum, the transcription factor SugR derived therefrom. The recombinant Corynebacterium glutamicum of the present invention increases the production of acetylglucosamine to up to 26 g/L, and lays a foundation for further metabolic engineering of Corynebacterium glutamicum to produce glucosamine.

Abstract: The present invention provides a method for promoting N-acetylglucosamine synthesis by using the GlcN6P responsive element. In the present invention, Bacillus subtilis BSGNY-Pveg-glmS-P43-GNA1 is used as a starting strain, in which a CRISPRi system regulated by GlcN6P responsive element is integrated into the genome to dynamically weaken the N-acetylglucosamine synthesis competitive pathway; a GlcN6P responsive promoter is used to regulate the expression of GNA1 on the plasmid to dynamically regulate the N-acetylglucosamine synthesis pathway; and the key gene alsSD involved in the acetoin synthesis pathway is knocked out. During fed-batch fermentation with this strain in a 15 L fermenter, the production of N-acetylglucosamine reaches 131.6 g/L and no by-product acetoin is accumulated, which lays a foundation for the production of GlcNAc by industrial fermentation.

Abstract: The present invention provides a pyruvate-responsive biosensor and a construction method and use thereof. In the present invention, pyruvate-responsive biosensors with different dynamic ranges are successfully constructed by optimizing the PdhR binding sequence inserted on the P43 promoter and optimizing the insertion site, wherein the minimum increase in dynamic range is by 0.6 time, and the maximum increase is by 30.7 times. The pyruvate-responsive biosensors are useful in the precise control of the expression of each gene in the cell. Since pyruvate is a key metabolite of central carbon in the cells, these biosensors are capable of dynamically regulating the expression level of intracellular genes according to changes in the content of pyruvate in the cells, thereby achieving the dynamic control of intracellular metabolic flux. The pyruvate biosensor obtained in the present invention has a good specificity, and a response range to pyruvate of 10-35 nmol/g DCW.

Abstract: The present invention provides an acid-tolerant Saccharomyces cerevisiae strain and use thereof. By using exogenously added malic acid as a stress, an acid-tolerant mutant S. cerevisiae strain MTPfo-4 is obtained by directed evolution screening in the laboratory, which tolerates a minimum pH of 2.44. The mutant strain MTPfo-4, tolerant to multiple organic acids, has an increased tolerance to exogenous malic acid of up to 86.6 g/L. The mutant strain MTPfo-4 obtained is further identified. The mutant strain grows stably and well, and can tolerate a variety of organic acids (lactic acid, malic acid, succinic acid, fumaric acid, citric acid, gluconic acid, and tartaric acid). It also has a strong tolerance to inorganic acids (HCl and H3PO4). This is difficult to achieve in the existing research and reports of S. cerevisiae. The strain is intended to be used as an acid-tolerant chassis cell factory for producing various short-chain organic acids.

Abstract: The present invention relates to a method for promoting acetylglucosamine synthesis of Bacillus subtilis, which belongs to the field of genetic engineering. The present invention adopts the recombinant Bacillus subtilis BSGNKAP2 as a starting strain, exogenously introducing pyruvate carboxylase BalpycA derived from Bacillus cereus, eliminating the central carbon metabolism overflow of the Bacillus subtilis and avoiding the synthesis of the by-product acetoin; further, five exogenous reducing force metabolic reactions are introduced to replace the reaction of generating NADH in glycolysis pathway and tricarboxylic acid cycle to reconstruct intracellular reducing force metabolism, which specifically comprise glyceraldehyde-3-phosphate ferredoxin dehydrogenase, isocitrate NAD+ dehydrogenase, a malate quinone dehydrogenase, a ketoacid ferredoxin oxidoreductase and a nitrogenase ferritin. In a shake-flask fermentation process using a complex medium, acetylglucosamine yield of the recombinant strain BSGNKAP8 is 24.

Abstract: The present invention provides a recombinant Bacillus subtilis with improved 2?-fucosyllactose production, and a construction method thereof. In the present invention, a strain capable of efficiently synthesizing 2?-fucosyllactose is obtained by the fusion expression of the fucosyltransferase gene and the L-fucokinase/guanosine 5?-diphosphate-L-fucose pyrophosphorylase gene in Bacillus subtilis BSGL-FF, the fermentation supernatant of which comprises a cumulative amount of 2?-fucosyllactose as high as 1.62 g/L, which is 55% higher than the amount achieved with the control strain. The construction method of the recombinant Bacillus subtilis of the present invention is simple, and convenient to use, and thus has good application prospects.

Abstract: The invention provides a recombinant Bacillus subtilis, construction method and use thereof, wherein the cell's own FMMs are used as a space scaffold, and a multi-enzyme complex is constructed from specific marker proteins FloA and FloT, such that an artificial substrate channel is formed, and the cell metabolic burden is effectively reduced. The recombinant Bacillus subtilis of the invention can efficiently synthesize GlcNAc without affecting cell life activity, and can also limit the toxic intermediate metabolite GlcN-6-P near the plasma membrane to reduce or eliminate its inhibition on cell activity. In the process of shaking flask fermentation using complex medium, the yield of acetyl glucosamine of the control strain BSG-C was only 0.45 g·L?1, while that of BSG-AT, BSG-ATP, BSG-ATPB increased to 5.29 g·L?1, 6.22 g·L?1 and 8.48 g·L?1 respectively. The construction method of recombinant Bacillus subtilis is simple, easy to use and has a good application prospect.

Abstract: The present disclosure provides a recombinant Bacillus subtilis for increasing the yield of menaquinone 7 (MK-7) and application thereof, and belongs to the field of genetic engineering. In the present disclosure, 14 recombinant strains BS1-BS14 are constructed through the modification of genes related to the biosynthetic pathway of MK-7 on a chromosome of Bacillus subtilis, wherein BS6-BS14 significantly increase the yield of the MK-7, reaching up to 33.5 mg/L, which is 3.53 times the yield of the original strain of wild-type Bacillus subtilis 168. The present disclosure further provides a method for modifying the MK-7 biosynthetic pathway in microorganisms to increase the yield of the MK-7, providing a theoretical basis for constructing a high-yielding strain of the MK-7.

Abstract: A port listening request dynamically generated by an application process hosted in a container can be identified. Whether the application process hosted in the container is trusted can be determined. Responsive to determining that the application process hosted in the container is trusted, a first port to be used as an external port for the application process can be dynamically selected, and a port assignment can be communicated to a container engine, the port assignment indicating the first port is assigned to the application process. The first port can be mapped to a second port assigned as an internal port for the application process. The first port can be opened for the application process.

Abstract: A port listening request dynamically generated by an application process hosted in a container can be identified. Whether the application process hosted in the container is trusted can be determined. Responsive to determining that the application process hosted in the container is trusted, a first port to be used as an external port for the application process can be dynamically selected, and a port assignment can be communicated to a container engine, the port assignment indicating the first port is assigned to the application process. The first port can be mapped to a second port assigned as an internal port for the application process. The first port can be opened for the application process.

Abstract: The disclosure discloses Bacillus subtilis for producing N-acetylneuraminic acid and application thereof, and belongs to the field of genetic engineering. The disclosure optimizes the expression levels of key enzymes in N-acetylneuraminic acid synthesis pathways on genome through promoters of different strength, reduces the protein synthesis pressure caused by the expression of enzymes on cells, and further integrates the three N-acetylneuraminic acids in a same Bacillus subtilis engineering strain. Bacillus subtilis with improved N-acetylneuraminic acid production is obtained, and the production reaches 10.4 g/L at the shake flask level, laying a foundation for further improving the NeuAc production from Bacillus subtilis.

Abstract: The disclosure herein relates to construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase, and belongs to the technical field of fermentation engineering. Firstly, recombinant B. subtilis capable of heterologously secreting and expressing a diacetylchitobiose deacetylase gene is constructed, and a signal peptide fragment yncM is added into the recombinant vector for the first time. The signal peptide can secrete the target protein diacetylchitobiose deacetylase outside the cells of the recombinant B. subtilis, and a mutant of the 5?-end untranslated region is acquired, thereby significantly increasing the expression level of the target protein, and greatly simplifying the subsequent enzyme separation and purification steps. When the acquired diacetylchitobiose deacetylase is fermented and cultured in a fermentation medium for 50-60 h, the enzyme activity reaches a maximum of 1,548.

Abstract: A computer system with programmable serial presence detection (SPD) data is disclosed. The computer system uses a user-programmable memory to store virtual SPD data which includes the configuration information of the memory module. The virtual SPD data is stored separately from the system boot code of the computer system. A computing unit uses a memory driver to drive the memory module. The memory driver accesses the user-programmable memory through a virtual SPD module to acquire the configuration information of the memory module.

Abstract: A computer system with programmable serial presence detection (SPD) data is disclosed. The computer system uses a user-programmable memory to store virtual SPD data which includes the configuration information of the memory module. The virtual SPD data is stored separately from the system boot code of the computer system. A computing unit uses a memory driver to drive the memory module. The memory driver accesses the user-programmable memory through a virtual SPD module to acquire the configuration information of the memory module.

Abstract: An extension lens module for an electronic device having a lens includes a seat body and a switching device. The seat body is configured to be detachably disposed on the electronic device and includes a positioning pillar. The switching device includes a rotating disc and a plurality of extension lenses, wherein the rotating disc is rotatable with respect to the seat body in an extending direction which is perpendicular to the seat body, and the extension lenses are disposed on the rotating disc at intervals.

Abstract: The disclosure herein relates to construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase, and belongs to the technical field of fermentation engineering. Firstly, recombinant B. subtilis capable of heterologously secreting and expressing a diacetylchitobiose deacetylase gene is constructed, and a signal peptide fragment yncM is added into the recombinant vector for the first time. The signal peptide can secrete the target protein diacetylchitobiose deacetylase outside the cells of the recombinant B. subtilis, and a mutant of the 5?-end untranslated region is acquired, thereby significantly increasing the expression level of the target protein, and greatly simplifying the subsequent enzyme separation and purification steps. When the acquired diacetylchitobiose deacetylase is fermented and cultured in a fermentation medium for 50-60 h, the enzyme activity reaches a maximum of 1,548.

Abstract: The invention relates to a recombinant strain of Bacillus subtilis, wherein pyruvate carboxylase BalpycA, glyceraldehyde-3-phosphate ferredoxin dehydrogenase gor, isocitrate NAD+ dehydrogenase icd, malate quinone dehydrogenase mqo, pyruvate ferredoxin oxidoreductase porAB and nitrogenase ferritin cyh are integrated and expressed in the recombinant strain. The invention also discloses use of the recombinant strain in fermentation production of acetylglucosamine. The recombinant Bacillus subtilis of the invention eliminates the central carbon metabolism overflow of the Bacillus subtilis and balances the intracellular reducing force, and the fermentation yield of acetylglucosamine is greatly improved.

Abstract: The invention discloses a promoter which can be induced to express in acidic conditions, and relates to the field of bioengineering technology. The promoters of the invention are separated from A. niger and can actuate and/or regulate the expression of the effectively connected nucleic acids in A. niger. In the invention the expression of the promoters is studied in A. niger, and it is indicated that some promoters show weak expression, and some show strong activity. The invention provides an effective method and new thought for organic acids production by fungi or other products produced by fermentation under acidic conditions.