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    • 1. 发明公开
    • 불용성 베타-1,3-글루칸을 분해하는 신규한 베타-글루카네이즈 및 그의 생산방법
    • 新型β-葡萄糖苷酶消化不溶性BET-1,3-GLUCAN及其方法
    • KR1020100034358A
    • 2010-04-01
    • KR1020080093446
    • 2008-09-23
    • 한국생명공학연구원네이처런스(주)
    • 최의성손여진이인영김미경박영일
    • C12N9/42C12N9/00C12Q1/68
    • C12N15/70C12N15/81
    • PURPOSE: A novel beta-glucanase is provided to ensure excellent hydrolysis activity to insoluble beta-glucan and produce soluble beta-glucan without chemical wastes. CONSTITUTION: A protein having an activity of beta-glucanase has an amino acid of sequence number 1 or 90% or more homology with the amino acid. A nucleic acid molecule encoding the beta-glucanase protein has nucleic acid of sequence number 2. The protein contains a beta-1,3-glucan binding domain. The protein is derived from Cellulomonas sp. A pBluescript KSII(+) vector contains the nucleic acid molecule encoding the beta-glucanase protein. The vector is a yeast expression vector and contains a promoter and secretion signal gene. The promoter is GAL10, GAPDH, PGK, ADH, PHO5, GAL1, SED1, MOX, TEF or TPI A gene encoding the secretion signal is MFα, PHO5, SUC2, AMY, SED or killer toxin. A transformant transformed by the vector is E. coli or Saccharomyces cerevisiae yeast. The beta-glucanase is obtained by culturing transformant and collecting beta-glucanase from culture.
    • 目的:提供一种新的β-葡聚糖酶,以确保对不溶性β-葡聚糖的优异的水解活性,并产生无化学废物的可溶性β-葡聚糖。 构成:具有β-葡聚糖酶活性的蛋白质具有与氨基酸序列1或90%以上同源性的氨基酸。 编码β-葡聚糖酶蛋白的核酸分子具有序列号2的核酸。蛋白质含有β-1,3-葡聚糖结合结构域。 蛋白质来源于纤维单胞菌属 pBluescript KSII(+)载体含有编码β-葡聚糖酶蛋白质的核酸分子。 载体是酵母表达载体,含有启动子和分泌信号基因。 启动子是GAL10,GAPDH,PGK,ADH,PHO5,GAL1,SED1,MOX,TEF或TPI编码分泌信号的基因是MFα,PHO5,SUC2,AMY,SED或杀伤毒素。 由载体转化的转化体是大肠杆菌或酿酒酵母(Saccharomyces cerevisiae)酵母。 通过培养转化体并从培养物中收集β-葡聚糖酶获得β-葡聚糖酶。
    • 7. 发明公开
    • 재조합단백질 생산용 단백질융합인자
    • 用于生产重组蛋白的翻译融合合作伙伴
    • KR1020070101190A
    • 2007-10-16
    • KR1020070091808
    • 2007-09-10
    • 한국생명공학연구원
    • 손정훈최의성배정훈윤남경성혜영정준기이홍원
    • C07K19/00
    • A novel translational fusion partner(TFP) is provided to express and secret a non-producible protein which is hard to be recombinantly produced. A TFP protein inducing the secretion of recombinant protein interleukin-2, colony stimulating factor(G-CSF) or lipase B(CalB) from eukaryotic cells is selected from the group consisting of (a) a TFP1 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 1 and an amino acid sequence having the TFP1 protein activity (b) a TFP2 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 3 and an amino acid sequence having the TFP2 protein activity (c) a TFP3 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 5 and an amino acid sequence having the TFP3 protein activity (d) a TFP4 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 7 and an amino acid sequence having the TFP4 protein activity (e) a TFP1-3 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 9 and an amino acid sequence having the TFP1-3 protein activity (f) a TFP1-4 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 10 and an amino acid sequence having the TFP1-4 protein activity (g) a TFP3-1-1 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 40 and an amino acid sequence having the TFP1-4 protein activity and (h) a TFP3-1-2 protein having 90-99.9% of homology to an amino acid sequence described as SEQ ID : NO. 42 and an amino acid sequence having the TFP3-1-2 protein activity.
    • 提供了一种新型翻译融合配偶体(TFP)来表达和分泌难以重组产生的非生产性蛋白质。 诱导来自真核细胞分泌重组蛋白质白细胞介素-2,集落刺激因子(G-CSF)或脂肪酶B(CalB)的TFP蛋白质选自(a)具有90-99.9%同源性的TFP1蛋白 涉及如SEQ ID NO:1所示的氨基酸序列。 1和具有TFP1蛋白活性的氨基酸序列(b)与SEQ ID NO:1所示的氨基酸序列具有90-99.9%同源性的TFP2蛋白。 3和具有TFP2蛋白活性的氨基酸序列(c)与SEQ ID NO:1所示的氨基酸序列具有90-99.9%同源性的TFP3蛋白。 5和具有TFP3蛋白活性的氨基酸序列(d)与SEQ ID NO:1所示的氨基酸序列具有90-99.9%同源性的TFP4蛋白。 7和具有TFP4蛋白活性的氨基酸序列(e)与SEQ ID NO:1所示的氨基酸序列具有90-99.9%同源性的TFP1-3蛋白。 9和具有TFP1-3蛋白活性的氨基酸序列(f)与SEQ ID NO:9所示氨基酸序列具有90-99.9%同源性的TFP1-4蛋白。 10和具有TFP1-4蛋白活性的氨基酸序列(g)TFP3-1-1蛋白质与SEQ ID NO:1所示的氨基酸序列具有90-99.9%的同源性。 40和具有TFP1-4蛋白活性的氨基酸序列和(h)TFP3-1-2蛋白质,其具有与SEQ ID NO:1所述的氨基酸序列具有90-99.9%的同源性。 42和具有TFP3-1-2蛋白活性的氨基酸序列。
    • 8. 发明公开
    • 신규한 베타-카로틴 케톨라제 및 베타-카로틴하이드록실라제 유전자
    • β-环糊精和β-硫酸羟脯氨酸的新基因编码
    • KR1020070077902A
    • 2007-07-30
    • KR1020060007749
    • 2006-01-25
    • 한국생명공학연구원
    • 최의성손정훈최은화손여진
    • C12N15/52
    • Novel genes for coding beta-carotene ketolase and beta-carotene hydroxylase are provided to be used for producing xanthopyll such as canthaxanthin, zeaxanthin and astaxanthin. The protein has the homology of more than 90% to an amino acid sequence described as SEQ ID : NO. 4, SEQ ID : NO. 8 or SEQ ID : NO. 6 and has the enzymatic activity of beta-carotene ketolase. The nucleic acid molecule codes the beta-carotene ketolase protein. The method for producing canthaxanthin comprises the steps of: (a) transforming a host cell using a vector coding the beta-carotene ketolase protein; and (b) culturing the transformant. The method for producing astaxanthin comprises the steps of: (a) transforming a host cell using a vector coding the beta-carotene ketolase protein and a vector coding the beta-carotene hydroxylase protein or using a vector coding the beta-carotene ketolase protein and the beta-carotene hydroxylase protein; and (b) culturing the transformant.
    • 提供用于编码β-胡萝卜素酮醇酶和β-胡萝卜素羟化酶的新基因用于生产黄体素如角黄素,玉米黄质和虾青素。 该蛋白质与SEQ ID NO:1所示的氨基酸序列具有超过90%的同源性。 4,SEQ ID NO: 8或SEQ ID:NO。 6,具有β-胡萝卜素酮醇酶的酶活性。 核酸分子编码β-胡萝卜素酮醇酶蛋白。 生产角黄素的方法包括以下步骤:(a)使用编码β-胡萝卜素酮醇酶蛋白的载体转化宿主细胞; 和(b)培养转化体。 制备虾青素的方法包括以下步骤:(a)使用编码β-胡萝卜素酮糖苷酶蛋白的载体和编码β-胡萝卜素羟化酶蛋白的载体或使用编码β-胡萝卜素酮醇酶蛋白的载体转化宿主细胞, β-胡萝卜素羟化酶蛋白; 和(b)培养转化体。
    • 10. 发明公开
    • 재조합단백질 생산용 단백질융합인자 라이브러리 및이로부터 획득된 단백질융합인자
    • 用于生产重组蛋白和翻译融合合作伙伴的翻译融合合作伙伴图书馆
    • KR1020070009269A
    • 2007-01-18
    • KR1020050064402
    • 2005-07-15
    • 한국생명공학연구원
    • 손정훈최의성배정훈신미경
    • C07K14/39C07K19/00C12N15/31
    • C12P21/02C07K14/39C12N15/1086C12N15/81
    • A method for selecting a translational fusion partner(TFP) is provided to be able to easily choose optimum TFP required for various target proteins by constructing a TFP library capable of inducing production of proteins, which are hard to be mass-produced due to low expression rate of yeast, from various gene resources. The method comprises the steps of: (a) preparing a variant cell where a reporter gene is deleted; (b) preparing an automatic selection vector where a reporter gene is inserted; (c) linking a gene fragment including a TFP inducing the secretion of a reporter protein with the automatic selection vector so as to prepare a TFP library(L); (d) transforming the library into a protein inactive cell to obtain an library from a cell showing the activity of reporter protein; (e) introducing the library obtained from the step(d) and a hard-to-express target protein gene(X) into a reporter-protein inactive cell so as to perform an in vivo homologous recombination; and (f) isolating a gene from the cell secreting the hard-to-express target protein and showing the reporter-protein activity and then analyzing characteristics of the TFP. In the method, the reporter gene is selected from the group consisting of invertase, amylase, glucoamylase, galactosidase, sucrase, cellulase, xylanase, maltase, phosphatase, beta-lactamase, and lipase and protease.
    • 提供了选择翻译融合配偶体(TFP)的方法,以便能够通过构建能够诱导蛋白质产生的TFP文库来容易地选择各种靶蛋白质所需的最佳TFP,由于低表达而难以大规模生产 来自各种基因资源的酵母菌率。 该方法包括以下步骤:(a)制备缺失报道基因的变体细胞; (b)制备插入报告基因的自动选择载体; (c)将包含诱导报告蛋白分泌的TFP的基因片段与自动选择载体连接以制备TFP文库(L); (d)将文库转化为蛋白质无活性细胞,从显示报告蛋白活性的细胞获得文库; (e)将从步骤(d)获得的文库和难以表达的靶蛋白基因(X)引入报告基因 - 蛋白质无活性的细胞中,以进行体内同源重组; 和(f)从分泌难以表达的靶蛋白的细胞中分离基因并显示报道蛋白活性,然后分析TFP的特征。 在该方法中,报告基因选自转化酶,淀粉酶,葡糖淀粉酶,半乳糖苷酶,蔗糖酶,纤维素酶,木聚糖酶,麦芽糖酶,磷酸酶,β-内酰胺酶,脂肪酶和蛋白酶。