The disclosure discloses a genetically engineered strain for producing porcine myoglobin and fermentation and purification thereof, and belongs to the technical field of genetic engineering. The disclosure realizes efficient secretion and expression of porcine myoglobin by integrating the gene of porcine myoglobin in P. pastoris. On this basis, optimization of the medium and culture conditions of recombinant P. pastoris can significantly increase the titer of porcine myoglobin, so that the titer can reach 285.42 mg/L under fermenter conditions. In addition, by creatively adding different concentrations of ammonium sulfate to fermentation broth step by step, the purity of myoglobin obtained by final concentration is up to 88.0%, and the purification rate is up to 66.1%. The disclosure realizes efficient expression and high purification of porcine myoglobin from various steps such as synthesis, fermentation and purification of porcine myoglobin, and provides broad prospects for industrial production of porcine myoglobin.

The present disclosure relates to a gene recombinant vector of a collagenase, comprising a collagenase gene, wherein an amino acid sequence of a collagenase encoded by the collagenase gene is shown in SEQ ID NO. 1; moreover, a genetically engineered strain of the collagenase and a preparation method of the collagenase are also disclosed; and the collagenase prepared according to the invention is capable of degrading a bone collagen, and improving a yield of a low-molecular-weight bone collagen peptide.

A tandem DNA element capable of enhancing protein synthesis efficiency, in particular, the nucleic acid construct is formed by an IRES enhancer (such as ScBOI1, ScFLO8, ScNCE102, ScMSN1, KlFLO8, KlNCE102, KlMSN1, KlBOI1) derived from eukaryotic cells (such as yeast), a Ω sequence, and a yeast-specific Kozak sequence in tandem. The use of the nucleic acid construct in a yeast-based in vitro biosynthesis system (such as a yeast-based in vitro protein synthesis system) can significantly improve protein synthesis efficiency.

Materials and methods that involve overexpression of a transcriptional activator such as retrograde regulation protein 1 (Rtg1) for increasing expression of one or more polypeptides.

The present disclosure application relates to a construction method of a yeast display library (YSD), specifically to a construct for a yeast display library, an expression vector, a host cell and a construction method and use thereof. The yeast display library provided by the present application has high transformation efficiency and rich diversity.

The objective of the present invention is to provide a transformed Aspergillus microorganism lacking at least two types of selection marker genes available for marker recycling method and a composition therefor. The objective can be achieved by a transformed Aspergillus microorganism lacking at least two types of selection marker genes available for marker recycling method on its chromosomes, or a composition for transforming an Aspergillus microorganism containing at least two types of nucleic acid fragments containing a loop-out region and a selection marker gene available for marker recycling method between homologous recombination regions, wherein the selection marker genes contain a tryptophan biosynthesis gene and a gene different from tryptophan biosynthesis gene.

The present invention relates to a leader peptide which promotes the secretion of recombinant proteins and a nucleic acid sequence encoding the leader peptide as well as expression cassettes, vectors and host cells comprising this leader sequence. Also disclosed is a method for producing a protein using this leader peptide.

The invention provides engineered biosynthetic pathways that can be used to produce cannabinoids from fatty acids, recombinant microorganisms incorporating such pathways, methods of biosynthetically producing cannabinoids from fatty acids, and cannabinoids so produced.

The present disclosure relates to a gene recombinant vector of a collagenase, comprising a collagenase gene, wherein an amino acid sequence of a collagenase encoded by the collagenase gene is shown in SEQ ID NO. 1; moreover, a genetically engineered strain of the collagenase and a preparation method of the collagenase are also disclosed; and the collagenase prepared according to the invention is capable of degrading a bone collagen, and improving a yield of a low-molecular-weight bone collagen peptide.

This application describes transcriptional units, synthetic expression systems, and host cells comprising transcriptional units and synthetic expression systems, wherein the synthetic expression system is capable of expressing a gene of interest. Also described are methods for the production of bioproducts (including, but not limited to, proteins or RNA expressed from the gene of interest). In some embodiments, bioproducts are produced from host cells under culture conditions without addition of methanol.