摘要:
A process for producing a lysosomal enzyme having a mannose-6-phosphate-containing acidic sugar chain, wherein the process comprising: culturing in a medium yeast cells obtained by introducing a lysosomal enzyme gene into a sugar chain biosynthetic enzyme gene mutant strain of yeast, collecting a lysosomal enzyme having a phosphate-containing sugar chain from the culture, and then treating the enzyme with α-mannosidase; and pharmaceutical compositions for treatment of human lysosomal enzyme deficiencies produced by the process. The genetic engineering technique using the yeast according to the present invention allows large-amount and high-purity production of a glycoprotein having a phosphate-containing acidic sugar chain which can serve as a labeling marker for transporting into lysosomes in cells of mammals such as human. The glycoprotein having a phosphate-containing acidic sugar chain according to the invention may be utilized as a drug effective in treatment of human lysosomal enzyme deficiencies, etc.
摘要:
A process for producing a lysosomal enzyme having a mannose-6-phosphate-containing acidic sugar chain, wherein the process comprising: culturing in a medium yeast cells obtained by introducing a lysosomal enzyme gene into a sugar chain biosynthetic enzyme gene mutant strain of yeast, collecting a lysosomal enzyme having a phosphate-containing sugar chain from the culture, and then treating the enzyme with α-mannosidase; and pharmaceutical compositions for treatment of human lysosomal enzyme deficiencies produced by the process. The genetic engineering technique using the yeast according to the present invention allows large-amount and high-purity production of a glycoprotein having a phosphate-containing acidic sugar chain which can serve as a labeling marker for transporting into lysosomes in cells of mammals such as human. The glycoprotein having a phosphate-containing acidic sugar chain according to the invention may be utilized as a drug effective in treatment of human lysosomal enzyme deficiencies, etc.
摘要:
This invention provides a means for high-level secretory production of a protein, and, in particular, a protein having a complicated structure such as an antibody, in a host cell such as a yeast cell. This invention provides a method for high-level secretory production of a foreign protein with the use of a transformed host cell having one or more types of chaperone protein genes and via suppression of O sugar chain inherent to a host cell such as a yeast cell.
摘要:
This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.
摘要:
This invention provides a means for enabling high-level secretory production of proteins, in particular proteins having complicated structures such as antibodies, in host cells such as yeast cells. The invention also provides transformed yeast cells having the activated HAC1 gene and the RRBP1 gene and a method for enabling high-level secretory production of foreign proteins using such transformed host cells by inhibiting O-sugar chain formation indigenous to host cells such as yeast cells.
摘要:
The present invention relates to a means for generating a mucin-type glycopeptide or glycoprotein on a large scale in yeast. Specifically, the invention relates to a method which comprises introducing into a yeast at least one selected from the group consisting of a gene encoding UDP-GalNAc synthetase, a gene encoding UDP-GalNAc transporter, and a gene encoding polypeptide:O-GalNAc transferase, and, if desired, a gene encoding a mucin-type glycopeptide; and producing a mucin-type glycoprotein having O-GalNAc by use of the yeast.
摘要:
This invention provides a means for enabling high-level secretory production of proteins, in particular proteins having complicated structures such as antibodies, in host cells such as yeast cells. The invention also provides transformed yeast cells having the activated HAC1 gene and the RRBP1 gene and a method for enabling high-level secretory production of foreign proteins using such transformed host cells by inhibiting O-sugar chain formation indigenous to host cells such as yeast cells.
摘要:
This invention provides a means for high-level secretory production of a protein, and, in particular, a protein having a complicated structure such as an antibody, in a host cell such as a yeast cell. This invention provides a method for high-level secretory production of a foreign protein with the use of a transformed host cell having one or more types of chaperone protein genes and via suppression of O sugar chain inherent to a host cell such as a yeast cell.
摘要:
A method for breeding yeast having thermotolerance or recovering growth activity and a method for breeding yeast which produces beta-glucan efficiently as well as an yeast obtained by such methods for breeding are presented by a method for breeding yeast having thermotolerance or recovering growth activity including a step for controlling proofreading function of DNA polymerase in a loss-of-function mutant of yeast (for example, a step for including mutant pol3 gene or mutant cdc6− gene in a gene-disruptant.
摘要:
A method for breeding yeast having thermotolerance or recovering growth activity and a method for breeding yeast which produces beta-glucan efficiently as well as an yeast obtained by such methods for breeding are presented by a method for breeding yeast having thermotolerance or recovering growth activity including a step for controlling proofreading function of DNA polymerase in a loss-of-function mutant of yeast (for example, a step for including mutant pol3 gene or mutant cdc6−gene in a gene-disruptant.