Abstract: The disclosure discloses recombinant Bacillus subtilis for synthesizing guanosine diphosphate fucose and a construction method and application thereof. The recombinant Bacillus subtilis is obtained by intensively expressing guanylate kinase and nucleotide diphosphokinase genes and expressing exogenous fucokinase and phosphate guanylyltransferase genes in a genome of Bacillus subtilis 168. According to the disclosure, a bacterial strain for synthesizing the guanosine diphosphate fucose is obtained by reconstructing the Bacillus subtilis 168, with a volume of intracellular accumulation up to 196.15 g/L. According to the disclosure, by intensively expressing the guanylate kinase and nucleotide diphosphokinase genes, and enhancing the supply of intracellular GDP-L-fucose composition cofactors, the synthesis of the guanosine diphosphate fucose is promoted. The construction method for the recombinant Bacillus subtilis of the disclosure is simple and convenient to use, thus having 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 disclosure provides an apparatus for testing spray and combustion performance of an internal-combustion engine based on a rapid compression-expansion machine, which belongs to the testing field of internal-combustion engines. It includes a driving mechanism, a transmission mechanism, an engine cylinder, an air intake system and an exhaust system. The engine cylinder includes a piston, a cylinder block, a dual-fuel cylinder head and a multi-injector assembly. The piston is disposed in the cylinder block. The driving mechanism is connected with the piston through the transmission mechanism and drives the piston to linearly move back and forth along a vertical direction of the cylinder block. The dual-fuel cylinder head is connected to an upper part of the cylinder block. The multi-fuel injection assembly includes a plurality of injectors. The plurality of injectors are of the same type and are all disposed on the dual-fuel cylinder head.

Abstract: The present disclosure provides a six-cylinder opposed free piston internal combustion engine generator. The generator comprises two free piston internal combustion engine sets, one opposed piston internal combustion engine set and two linear generator sets. Air entering cylinders is subjected to first-stage compression in low-pressure cylinder sets in the free piston internal combustion engine sets and the opposed piston internal combustion engine set and then subjected to second-stage compression in high-pressure cylinder sets, and a high pressure gas produced after the combustion is subjected to first-stage expansion in the high-pressure cylinder sets and then subjected to second-stage expansion in the low-pressure cylinder sets.

Abstract: The present disclosure provides an apparatus for testing spray and combustion performance of an internal-combustion engine based on a rapid compression-expansion machine, which belongs to the testing field of internal-combustion engines. The present disclosure solves the problems that the existing test of the internal-combustion engine testing apparatus having a complex structure, is poor in adaptability, cannot be observed easily, and cannot meet the requirements of multi-fuel injection and multi-injection-pressure tests. It includes a driving mechanism, a transmission mechanism, an engine cylinder, an air intake system and an exhaust system. The engine cylinder includes a piston, a cylinder block, a dual-fuel cylinder head and a multi-injector assembly. The piston is disposed in the cylinder block. The driving mechanism is connected with the piston through the transmission mechanism and drives the piston to linearly move back and forth along a vertical direction of the cylinder block.

Abstract: The disclosure is an engine simulation test device capable of realizing an ultrahigh compression temperature and pressure, belonging to the field of a diesel engine high-temperature and high-pressure system, solving the problems that the existing engine simulation test cannot achieve ultra-high compression temperature and pressure, and the temperature and pressure are not adjustable. It includes a compressed air inlet mechanism, a nitrogen gas inlet mechanism, a pressure stabilizing mechanism, a cyclic heating mechanism, an air inlet mechanism and a fast compressor mechanism.

Abstract: A free piston generator based on a rigid synchronous transmission system is provided, which belongs to the technical field of power energy. The present disclosure solves the problems of low power generation efficiency and low stability of the existing free piston generator. The free piston generator based on the rigid synchronous transmission system includes a first linear generator set, a second linear generator set, a rigid synchronous transmission assembly, two high-pressure cylinders arranged at two ends of the first linear generator set, and two low-pressure cylinders arranged at two ends of the second linear generator set.

Abstract: Provided is a free piston generator based on a split thermodynamic cycle, which belongs to the technical field of power energy. The present disclosure solves the problem of low power generation efficiency of an existing free piston generator. The free piston generator includes a linear generator set and two internal combustion engine sets arranged at two ends of the linear generator set. Air is first subjected to first-stage compression by the low-pressure cylinder set in the internal combustion engine sets and is then subjected to second-stage compression in the high-pressure cylinders, so that the intake pressure of an internal combustion engine is effectively increased, which is favorable for increasing the average effective pressure in a work process, thereby improving the thermal efficiency and the power generation efficiency of the free piston generator.

Abstract: The present disclosure provides a six-cylinder opposed free piston internal combustion engine generator. The generator comprises two free piston internal combustion engine sets, one opposed piston internal combustion engine set and two linear generator sets. Air entering cylinders is subjected to first-stage compression in low-pressure cylinder sets in the free piston internal combustion engine sets and the opposed piston internal combustion engine set and then subjected to second-stage compression in high-pressure cylinder sets, and a high pressure gas produced after the combustion is subjected to first-stage expansion in the high-pressure cylinder sets and then subjected to second-stage expansion in the low-pressure cylinder sets.

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.