Provided herein are compositions with enhanced protein content, protein compositions with improved functionality, and methods for the preparation thereof.

The disclosure discloses the preparation of thermophilic -glucosidase and the application thereof, which belongs to the technical field of genetic engineering and fermentation engineering. The present disclosure heterologously expresses the -glucosidase TpBgl3A derived from thermophilic fungus Talaromyces piceae by constructing a recombinant bacteria. The enzyme production of recombinant bacteria can reach 2324 U/mL in a 3.6 L fermenter. The resulting -glucosidase TpBgl3A can produce gentioligosaccharides with a high conversion rate using glucose as a substrate at a lower enzyme amount added, which significantly reduces production costs. In the reaction system using glucose and cellobiose as substrates, the conversion rate of gentioligosaccharide reaches 26.2%, which has the potential for industrial production of gentioligosaccharide.

The present invention provides polynucleotide vectors for high expression of heterologous genes. Some vectors further comprise novel transposons and transposases that further improve expression. Further disclosed are vectors that can be used in a gene transfer system for stably introducing nucleic acids into the DNA of a cell. The gene transfer systems can be used in methods, for example, gene expression, bioprocessing, gene therapy, insertional mutagenesis, or gene discovery.

The invention discloses a fusion protein and its preparation method for intermediate polypeptide of Semaglutide. The invention belongs to the technical field of genetic engineering and polypeptide preparation. The fusion protein comprises a fusion peptide, a protease cleavage site and a target main molecular sequence. By optimizing the fusion peptide sequence, changing the isoelectric point and hydrophilicity of the protein, the highest expression of the fusion protein is effectively increased to 13.1 g/L. Meanwhile, the properties of fusion protein are also improved, which is conducive to the development of subsequent extraction, enzyme digestion and purification processes. The yield of intermediate polypeptide after enzyme digestion is 3.62 g/L. The production cost of Semaglutide intermediate polypeptide Arg34GLP-1(9-37) is reduced from the source, which is conducive to industrial scale-up and suitable for industrial production.

A novel method of growing fungi is disclosed which uses an engineered artificial media and produces high density filamentous fungi biomats that can be harvested with a minimum of processing and from which fungal products such as antibiotics, proteins, and lipids can be isolated, the method resulting in lowered fungus cultivation costs for energy usage, oxygenation, water usage and waste stream production.

A high temperature resistant mannanase mutant, comprising at least one mutation site among D49C/I/N, G58D/E/S, H90L, T99Y, K120Y, N124P, T130D/N/P/Q/R/S, Y132F, S136M, S140L/K, T148K, F206Y, A212N/S/P, L257F, S263R, D268A/E/G/P, G273P, S302M/E, S306M/Q, T315P, T353L/F, D360E/N/K, and A362R. The high temperature resistant mannanase mutant has heat resistance significantly higher than the wild type mannanase, and can be widely used in the field of feeds.

A method for producing a recombinant collagen-like protein (CLP) can be performed. The method includes fermenting a host cell, accumulating the CLP in a medium to obtain a fermentation broth, separating the host cell from the fermentation broth to obtain a supernatant, and incubating the supernatant. The CLP can be purified after incubation.

Disclosed herein are genetically engineered yeast cells capable of producing lactate from sucrose. The genetically engineered yeast cells comprise a polynucleotide encoding an exogenous lactate dehydrogenase enzyme: a polynucleotide encoding an exogenous invertase enzyme: a deletion or disruption of a native pyruvate decarboxylase (PDC) gene: and a genetic modification resulting in overexpression of a native hexokinase gene.

A novel method of growing fungi is disclosed which uses an engineered artificial media and produces high density filamentous fungi biomats that can be harvested with a minimum of processing and from which fungal products such as antibiotics, proteins, and lipids can be isolated, the method resulting in lowered fungus cultivation costs for energy usage, oxygenation, water usage and waste stream production.

Several mutated strains of Scheffersomyces stipitis are generated by repetitive culturing of the parent strain on two types of concentrated hydrolyzates and with ethanol-challenged xylose-fed continuous culture. Isolates collected from various enriched populations are screened and ranked based on relative xylose uptake rate and ethanol yield. Ranking on hydrolyzates with and without nutritional supplementation is used to identify those isolates with best performance across diverse conditions.