Provided is a hepatitis B treatment vaccine based on an inactivated whole recombinant Hansenula polymorpha cell which expresses HBsAg and HBcAg. An HBsAgVLP and an HBcAgVLP expressed in the recombinant Hansenula polymorpha cell are used as antigens, the amino acid sequence of the HBsAg expressed by the recombinant Hansenula polymorpha contains a total of 19 CTL epitopes, the amino acid sequence of the HBcAg expressed by the recombinant Hansenula polymorpha contains a total of 19 CTL epitopes, and the inactivated whole recombinant Hansenula polymorpha cell is used as an adjuvant.

Strains of yeast genetically engineered to produce increased amounts of non-hydroxylated collagen or hydroxylated collagen are described. An all-in-one vector including the DNA necessary to produce collagen, promotors, and hydroxylating enzymes is also described. Methods for producing non-hydroxylated or hydroxylated collagen are also provided.

Disclosed is a method for manufacturing a monoclonal antibody without using animal individuals. This method includes a step of introducing a DNA fragment comprising a gene that encodes a secretory signal, a gene that encodes a nanobody, and a gene that encodes a peptide barcode, or a vector containing the DNA fragment, into a yeast cell; and a step of collecting a polypeptide comprising the nanobody and the peptide barcode that has been expressed in the cell and secreted to the outside of the cell. According to the method, it is possible to manufacture a monoclonal nanobody more efficiently in a shorter period of time without using animal individuals.

Pichia pastoris alcohol dehydrogenase 2 (ADH2) promoter variants include at least one of the specified modifications on wild-type Pichia pastoris ADH2 promoter (SEQ ID NO: 1). The modification includes one of the following mutations: integration of a Cat8 transcription factor binding site (TFBS), particularly integration of SEQ ID NO: 3 or other gene sequences that show at least 80% similarity with this sequence, at any positions within nucleotides a) 647 to 660; b) 739 to 752; c) 1 to 948; and d) mutations specified with SEQ ID NO: 2 within nucleotides 15 to 848 separately and combinations thereof.

Provided is a hepatitis B treatment vaccine based on an inactivated whole recombinant Hansenula polymorpha cell which expresses HBsAg and HBcAg. An HBsAgVLP and an HBcAgVLP expressed in the recombinant Hansenula polymorpha cell are used as antigens, the amino acid sequence of the HBsAg expressed by the recombinant Hansenula polymorpha contains a total of 19 CTL epitopes, the amino acid sequence of the HBcAg expressed by the recombinant Hansenula polymorpha contains a total of 19 CTL epitopes, and the inactivated whole recombinant Hansenula polymorpha cell is used as an adjuvant.

A recombinant bacterium for producing L-lysine, a construction method thereof, and a method for producing L-lysine by using the recombinant bacterium. The recombinant bacterium has increased expression and/or activity of asparaginase compared to a starting bacterium.

Provided is a recombinant Hansenula polymorpha-based high dosage hepatitis B vaccine, an HBsAg pure stock solution yield of a recombinant Hansenula polymorpha fermentation broth used for producing the hepatitis B vaccine being 300 mg/L-400 mg/L.

A hepatitis B treatment vaccine on the basis of inactivated, whole recombinant Hansenula polymorpha cells expressing HBsAg. The vaccine is the HBsAg expressed in recombinant Hansenula polymorpha cells. 10.sup.8 cells contain 6-10 g HBsAg as an antigen; the vaccine contains a total of 16-21 HBsAg-specific CTL epitopes; the vaccine uses optimized inactivated, fully recombinant Hansenula polymorpha cells as an adjuvant.

Provided is an engineered bacterium for producing nervonic acid and/or grease. The genome of the engineering bacterium is integrated with an expression cassette expressing a protein encoded by 3-ketoacyl-CoA synthase (KCS) gene and/or an exterase gene.

A hepatitis B treatment vaccine on the basis of inactivated, fully recombinant Hansenula cells expressing HBsAg. The vaccine is the HBsAg expressed in recombinant Hansenula cells. 10.sup.8 cells contain 6-10 g HBsAg as an antigen; the vaccine contains a total of 16-21 HBsAg-specific CTL epitopes; the vaccine uses optimized inactivated, fully recombinant Hansenula cells as an adjuvant.