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1. 发明授权

US07897724B2 Solid phase Fmoc chemistry process to prepare peptides 失效
标题翻译：固相Fmoc化学过程制备肽
公开(公告)号：US07897724B2
公开(公告)日：2011-03-01
申请号：US11729047
申请日：2007-03-27
申请人： Divya Lal Saksena , Shrikant Mishra , Chandrakesan Muralidharan , Nilesh Patil , Nikhil Umesh Mohe , Mandar Ravindra Maduskar
发明人： Divya Lal Saksena , Shrikant Mishra , Chandrakesan Muralidharan , Nilesh Patil , Nikhil Umesh Mohe , Mandar Ravindra Maduskar
IPC分类号： C07K1/06
CPC分类号： C07K14/75 , C07K14/78 , Y02P20/582
摘要： The present invention relates to an improved process for the preparation of N6-(aminoiminomethyl)-N2-(3-mercapto-1-oxopropyl)-L-lysylglycyl-L-α-aspartyl-L-tryptophyl-L-prolyl-L-cysteinamide, cyclic(1→6)-disulfide of formula (1), which involves assembling a peptide chain comprising of six amino acids and a thioalkyl carboxylic acid in a required sequence on a solid support to obtain a peptide bound resin of formula (2), capping the free amino groups after each coupling, cleaving Dde group in the peptide of formula (2) from the solid support to obtain peptide-solid support of formula (3), guanylating the peptide of formula (3) at ε-lysine-NH2 in an organic solvent to obtain peptide-solid support of formula (4), cleaving and deprotecting all groups in the peptide of formula (4) from the solid support to obtain peptide-amide formula (5), oxidizing the SH-peptide of formula (5) with an appropriate oxidizing agent to obtain the crude peptide-amide of formula (1) and purifying the crude peptide-amide of formula (1) by chromatographic technique. The solid support is either resin or a cellulose support like cotton, gauze, fabric, paper and perloza beads. The described process is simple, easy, environment friendly, takes lesser time and more cost effective.
摘要翻译：本发明涉及一种制备N6-（氨基亚氨基甲基）-N2-（3-巯基-1-氧代丙基）-L-赖氨酰甘氨酰-L-α-天冬氨酰-L-色氨酰-L-脯氨酰基-L- 半胱氨酰胺，式（1）的环状（1→6） - 二硫化物，其包括在固体支持物上以所需的顺序将包含六个氨基酸的肽链和硫代烷基羧酸组装以获得式（2）的肽结合树脂），在每次偶联后封端游离氨基，从固体支持物切割式（2）的肽中的Dde基团以获得式（3）的肽 - 固体支持物，在式（3）的肽上进行脒化，赖氨酸-NH 2在有机溶剂中得到式（4）的肽固体支持物，从固体支持物中切割和脱保护基团中所有基团的肽，得到肽 - 酰胺式（5），氧化SH- 式（5）的肽与适当的氧化剂反应，得到式（1）的粗肽 - 酰胺并纯化粗蛋白 e通过色谱技术制备式（1）的肽 - 酰胺。固体支持物是树脂或纤维素载体，如棉，纱布，织物，纸和perloza珠。描述的过程简单，容易，环保，花费更少的时间和更具成本效益。

2. 发明申请

US20070249806A1 Solid phase Fmoc chemistry process to prepare peptides 失效
标题翻译：固相Fmoc化学过程制备肽
公开(公告)号：US20070249806A1
公开(公告)日：2007-10-25
申请号：US11729047
申请日：2007-03-27
申请人： Divya Saksena , Shrikant Mishra , Chandrakesan Muralidharan , Nilesh Patil , Nikhil Mohe , Mandar Maduskar
发明人： Divya Saksena , Shrikant Mishra , Chandrakesan Muralidharan , Nilesh Patil , Nikhil Mohe , Mandar Maduskar
IPC分类号： C07K7/60 , C07K7/06
CPC分类号： C07K14/75 , C07K14/78 , Y02P20/582
摘要： The present invention relates to an improved process for the preparation of N6-(aminoiminomethyl)-N2-(3-mercapto-1-oxopropyl)-L-lysylglycyl-L-α-aspartyl-L-tryptophyl-L-prolyl-L-cysteinamide, cyclic(16)-disulfide of formula (1), which involves assembling a peptide chain comprising of six amino acids and a thioalkyl carboxylic acid in a required sequence on a solid support to obtain a peptide bound resin of formula (2), capping the free amino groups after each coupling, cleaving Dde group in the peptide of formula (2) from the solid support to obtain peptide-solid support of formula (3), guanylating the peptide of formula (3) at ε-lysine-NH2 in an organic solvent to obtain peptide-solid support of formula (4), cleaving and deprotecting all groups in the peptide of formula (4) from the solid support to obtain peptide-amide formula (5), oxidizing the SH-peptide of formula (5) with an appropriate oxidizing agent to obtain the crude peptide-amide of formula (1) and purifying the crude peptide-amide of formula (1) by chromatographic technique. The solid support is either resin or a cellulose support like cotton, gauze, fabric, paper and perloza beads. The described process is simple, easy, environment friendly, takes lesser time and more cost effective
摘要翻译：本发明涉及一种制备N 6 - （氨基亚氨基甲基）-N 2 - （3-巯基-1-氧代丙基）-L-赖氨酰甘氨酰基 - L-α-天冬氨酰基-L-色氨酰-L-脯氨酰基-L-半胱氨酰胺，式（1）的环状（16） - 二硫化物，其包括以所需顺序组装包含六个氨基酸的肽链和硫代烷基羧酸在固体支持物上获得式（2）的肽结合树脂，每次偶联后封端游离氨基，从固体支持物切割式（2）的肽中的Dde基团以获得式（3）的肽 - 固体支持物），在有机溶剂中在ε-赖氨酸-NH 2上对式（3）的肽进行脒化，得到式（4）的肽固体支持物，将式（4）的肽中的所有基团切割和脱保护（4）从固体支持物得到肽 - 酰胺配方（5），用合适的氧化剂氧化式（5）的SH-肽，得到粗肽 - 酰胺通过色谱技术纯化式（1）的粗肽酰胺。固体支持物是树脂或纤维素载体，如棉，纱布，织物，纸和perloza珠。描述的过程简单，容易，环保，花费更少的时间和更具成本效益

3. 发明授权

US08298789B2 Orthogonal process for purification of recombinant human parathyroid hormone (rhPTH) (1-34) 有权
标题翻译：纯化重组人甲状旁腺激素（rhPTH）（1-34）的正交方法
公开(公告)号：US08298789B2
公开(公告)日：2012-10-30
申请号：US12452919
申请日：2007-12-06
申请人： Uma Sankararaman , Dipanwita Maiti , Meera Sankarankutty , Rakesh Shekhawat , Narasimha Kumar Kopparapu , Gulnaz Zaidi , Bipinchandra Rathod , Niren Praful Thakar , Priti Thakur , Anjali Chutke , Shrikant Mishra
发明人： Uma Sankararaman , Dipanwita Maiti , Meera Sankarankutty , Rakesh Shekhawat , Narasimha Kumar Kopparapu , Gulnaz Zaidi , Bipinchandra Rathod , Niren Praful Thakar , Priti Thakur , Anjali Chutke , Shrikant Mishra
IPC分类号： C12N15/16
CPC分类号： C07K14/635 , C07K2319/21 , C07K2319/35 , C07K2319/50 , C07K2319/61
摘要： The present invention discloses a process for the preparation of rhPTH (1-34) also known as teriparatide by construction of a novel nucleotide, as an NcoI.IXhoI fragment as set forth in SEQ. ID. No.:1 encoding a chimeric fusion protein as set forth in SEQ.ID. No.:2 comprising of a fusion partner consisting of 41 amino acids belonging to Escherichia coli β-galactosidase (LacZ) gene, an endopeptidase cleavage site, rhPTH (1-34) gene fragment, cloning the said nucleotide in an expression vector under the control of T7 promoter, transforming Escherichia coli with the said vector and expressing the chimeric fusion protein in fed batch fermentation. The present invention further discloses a low feed rate lactose induction for optimized expression of rhPTH (1-34) in Escherichia coli. The present invention also discloses an unique, novel two step orthogonal purification process for rhPTH (1-34) comprising of cation exchange chromatography optionally followed by preparative chromatography selected from HIC or RP-HPLC to yield a target protein of ≧99% purity. The present invention discloses a simple, cost-effective, environmentally benign method of producing high purity rhPTH (1-34).
摘要翻译：本发明公开了一种制备rhPTH（1-34）的方法，其也称为特立帕肽，通过构建新的核苷酸，如SEQ ID NO：1所示的NcoI.IXhoI片段。 ID。编号如SEQ.ID所示的嵌合融合蛋白。编号：2包含由属于大肠杆菌和半乳糖苷酶（LacZ）基因的41个氨基酸组成的融合配偶体，内肽酶切割位点，rhPTH（1-34）基因片段，将所述核苷酸克隆在表达载体中 T7启动子的控制，用所述载体转化大肠杆菌并在补料分批发酵中表达嵌合融合蛋白。本发明还公开了用于在大肠杆菌中优化rhPTH（1-34）表达的低进料速率乳糖诱导。本发明还公开了一种用于rhPTH（1-34）的独特的新型两步正交纯化方法，其包括阳离子交换色谱，任选地随后进行选自HIC或RP-HPLC的制备型色谱，以产生≥99％纯度的靶蛋白。本发明公开了一种生产高纯度rhPTH（1-34）的简单，成本效益高，环境友好的方法。

4. 发明申请

US20100047870A1 LOW CELL DENSITY FERMENTATION PROCESS FOR THE PRODUCTION OF HETEROLOGOUS RECOMBINANT PROTEINS IN MICROORGANISMS 审中-公开
标题翻译：用于生产微生物中异位重组蛋白的低密度发酵方法
公开(公告)号：US20100047870A1
公开(公告)日：2010-02-25
申请号：US11722504
申请日：2004-12-21
申请人： Milind Prabhakar Niphadkar , Genevieve Suzle Nazareth , Neelesh Ramesh Surlikar , Monsur Ahmed Borbhuiya , Uma Sankararaman , Dipanwita Maiti , Laxmi Srinivas Rao , Saptarshi Paul , Shrikant Mishra
发明人： Milind Prabhakar Niphadkar , Genevieve Suzle Nazareth , Neelesh Ramesh Surlikar , Monsur Ahmed Borbhuiya , Uma Sankararaman , Dipanwita Maiti , Laxmi Srinivas Rao , Saptarshi Paul , Shrikant Mishra
IPC分类号： C12P21/00 , C12N9/38
CPC分类号： C12P21/02 , C12N1/20
摘要： A low cell density fermentation process for the production of heterologous proteins in microorganisms. The cell culture obtained by cultivating host microorganisms transformed with a vector carrying genetic material for the said proteins and an inducible promoter under batch fermentation conditions is fed with a feed medium after an OD600 of 0.16 to 8 has been achieved or after 0 to 4 hrs from the start of the fermentation process. The feed medium comprises 5 to 30% of carbon source and 1 to 30% of nitrogen source and 0 mg to 400 mg antibiotics and 2.5 to 4.25% inorganic phosphates and trace elements. The concentration of the carbon source in the feed medium is 10 to 30 and the amino acid content in nitrogen source is 45 to 95%. The initial feed rate is in the range of 0 ml/hr to 12 ml/hr and is raised exponentially by an exponent in the range of 0.1 to 0.4 and/or linearly with the slope of the curve in the range of 0.5 to 3. The production of the heterologous proteins is induced with 0.01-4% inducer at a cell density of OD600 0.1-OD600 50. The feeding of the cell culture with the feed medium and the feed rate described above is continued after production has been induced. The pO2 is adjusted between 10% to 60% by passing sterile air into the fermentation broth and the temperature and pH of the fermentation broth are maintained at 33° C.-41° C. and 6.9 - 8.5, respectively during the entire fermentation.
摘要翻译：用于在微生物中产生异源蛋白质的低细胞密度发酵方法。在分批发酵条件下培养用携带所述蛋白质的遗传物质的载体转化的宿主微生物获得的细胞培养物在实现了OD600为0.16至8或0至4小时之后的饲料培养基中进料开始发酵过程。饲料培养基含有5-30％的碳源和1-10％的氮源和0mg-400mg的抗生素和2.5-4.25％的无机磷酸盐和微量元素。饲料培养基中碳源的浓度为10〜30，氮源中氨基酸含量为45〜95％。初始进料速率在0毫升/小时至12毫升/小时的范围内，并且在0.1至0.4范围内以指数指数增加和/或以0.5至3的范围内曲线的斜率线性地升高。以OD 600 0.1-OD 60050的细胞密度用0.01-4％诱导剂诱导异源蛋白的产生。在诱导生产后，继续用饲料培养基喂养细胞培养物和上述进料速率。通过将无菌空气送入发酵液中，将pO2调节在10％至60％之间，发酵液的温度和pH分别在整个发酵期间保持在33℃-41℃和6.9-8.5℃。

5. 发明申请

US20080182781A1 Process for the Preparation of Peptides 审中-公开
标题翻译：肽的制备方法
公开(公告)号：US20080182781A1
公开(公告)日：2008-07-31
申请号：US10592118
申请日：2004-10-10
申请人： Divya Saksena , Shrikant Mishra , Chandrakesan Muralidharan , Milind Thakare , Nilesh Patil , Aruna Khare
发明人： Divya Saksena , Shrikant Mishra , Chandrakesan Muralidharan , Milind Thakare , Nilesh Patil , Aruna Khare
IPC分类号： A61K38/12 , C07K7/64 , C07K7/06
CPC分类号： C07K7/06 , Y02P20/55
摘要： The present invention relates to an improved process for the preparation of N6-(aminoiminomethyl)-N2-(3-mercapto-1-oxopropyl-L-lysylglycyl-L-α-aspartyl-L-tryptophyl-L-prolyl-L-cysteinamide, cyclic(1→6)-disulfide of formula (1), which involves assembling amino acid residues and a thioalkyl carboxylic acid with appropriate protecting groups on a solid phase resin, cleaving the peptide thus obtained from the resin with concomitant removal of side chain protecting groups except Acm protecting group of thiol moiety to obtain peptide amide of formula (3), converting lysine residue of peptide amide of formula (3) having protected thiol group to homoarginine residue by guanylation to obtain peptide of formula (4), preparing silver peptide of formula (5), followed by simultaneous deprotection, obtaining silver peptide of formula (5) and oxidation of silver peptide to obtain crude peptide amide comprising compound of formula (1) and finally subjecting to chromatographic purification. The described process is simple, easy, environment friendly and cost effective.
摘要翻译：本发明涉及一种制备N 6 - （氨基亚氨基甲基）-N 2 - （3-巯基-1-氧代丙基-L-赖氨酰甘氨酰-L- -α-天冬氨酰基-L-色氨酰-L-脯氨酰基-L-半胱氨酰胺，式（1）的环状（1-> 6） - 二硫化物，其包括将氨基酸残基和硫代烷基羧酸与固体上适当的保护基团组装通过除去除巯基部分的Acm保护基之外的侧链保护基除去由树脂得到的肽，从而得到式（3）的肽酰胺，转化具有被保护的硫醇的式（3）的肽酰胺的赖氨酸残基通过胍基化得到高精氨酸残基，得到式（4）的肽，制备式（5）的银肽，然后同时去保护，得到式（5）的银肽和银肽的氧化，得到粗肽酰胺，式（1），最后进行色谱 c净化。描述的过程简单，容易，环保，成本效益高。

6. 发明申请

US20090239262A1 Affinity Polypeptide for Purification of Recombinant Proteins 审中-公开
标题翻译：亲和多肽用于纯化重组蛋白
公开(公告)号：US20090239262A1
公开(公告)日：2009-09-24
申请号：US12161603
申请日：2007-02-07
申请人： Laxmi Srinivas Rao , Shrikant Mishra , Monsur Ahmed Borbhuiya , Deepa Mhatre , Priti Thakur
发明人： Laxmi Srinivas Rao , Shrikant Mishra , Monsur Ahmed Borbhuiya , Deepa Mhatre , Priti Thakur
IPC分类号： C12P21/02 , C07K14/00 , C12N15/11 , C12N15/00 , C12N1/21 , C12N1/19 , C12N5/06
CPC分类号： C12N15/62
摘要： The present disclosure provides an affinity polypeptide for the purification of a recombinant biologically active protein or polypeptide. Further, the present disclosure provides a fusion recombinant protein or polypeptide wherein the fusion recombinant protein comprises of at least two components, a biologically active polypeptide or protein or protein of interest and the affinity polypeptide. The biologically active polypeptide may be linked directly or indirectly to the affinity polypeptide by covalent binding. The present disclosure provides a recombinant expression vector for the producing said fusion recombinant protein in host cells. Further, the present disclosure provides an improved method of purification of recombinant protein from the host cells. Further, the disclosure provides a method of purification of the recombinant biologically active polypeptide or protein by immobilized metal ion chelating chromatography.
摘要翻译：本公开提供了用于纯化重组生物活性蛋白或多肽的亲和多肽。此外，本公开提供了融合重组蛋白或多肽，其中融合重组蛋白包含至少两种组分，生物活性多肽或感兴趣的蛋白质或亲和性多肽。生物活性多肽可以通过共价结合直接或间接地连接到亲和多肽上。本公开内容提供了用于在宿主细胞中产生所述融合重组蛋白的重组表达载体。此外，本公开提供了从宿主细胞纯化重组蛋白质的改进方法。此外，本公开提供了通过固定金属离子螯合色谱法纯化重组生物活性多肽或蛋白质的方法。

7. 发明申请

US20090035815A1 Synthetic Gene for Enhanced Expression in E. Coli 审中-公开
标题翻译：在大肠杆菌中增强表达的合成基因
公开(公告)号：US20090035815A1
公开(公告)日：2009-02-05
申请号：US12126319
申请日：2008-05-23
申请人： Laxmi Srinivas Rao , Monsur Borbhuiya Ahmed , Milind Niphadkar , Dinesh Paliwal , Rakesh Shekhawat , Aruna Khare , Radhika Samant , Priti Thakur , Anjali Chutke , Neelesh Surlikar , Sagar Zawar , Shrikant Mishra
发明人： Laxmi Srinivas Rao , Monsur Borbhuiya Ahmed , Milind Niphadkar , Dinesh Paliwal , Rakesh Shekhawat , Aruna Khare , Radhika Samant , Priti Thakur , Anjali Chutke , Neelesh Surlikar , Sagar Zawar , Shrikant Mishra
IPC分类号： C12P21/06 , C12N15/62 , C12P41/00 , C07H21/04 , C12N15/12
CPC分类号： C12P21/02 , C07K14/58
摘要： A novel nesiritide synthetic cDNA chimera encoding human b-type natriuretic peptide (hBNP) or nesiritide and a process for the preparation of the said novel chimera. Further, the inventors disclose the use of nesiritide synthetic cDNA chimera to obtain an expressible construct to produce mature nesiritide. Particularly, the inventors disclose an application of recombinant cloning method to prepare an ORF of a nesiritide chimeric construct, which is simultaneously codon optimized for E. coli and has optimal RNA stability. The inventors also provide a process for large scale purification of nesiritide by pH precipitation and chromatography.
摘要翻译：编码人b型利尿钠肽（hBNP）或奈西立肽的新型奈西立肽合成cDNA嵌合体及其制备方法。此外，发明人公开了使用奈西立肽合成cDNA嵌合体以获得可表达的构建体以产生成熟的奈西立肽。特别地，本发明人公开了重组克隆方法的应用，以制备奈西立肽嵌合构建体的ORF，其同时对大肠杆菌进行密码子优化并具有最佳的RNA稳定性。本发明人还提供了通过pH沉淀和色谱法大规模纯化奈西立胺的方法。

8. 发明申请

US20080167452A1 Process for Purification of Human Growth Hormone 审中-公开
标题翻译：纯化人类生长激素的方法
公开(公告)号：US20080167452A1
公开(公告)日：2008-07-10
申请号：US11568235
申请日：2004-11-09
申请人： Dipanwita Maiti , Shrikant Mishra , Laxmi Srinivas Rao , Milind Prabhakar Niphadkar , Ahmed Monsur Borbhuiya , Madhava Yada Rao
发明人： Dipanwita Maiti , Shrikant Mishra , Laxmi Srinivas Rao , Milind Prabhakar Niphadkar , Ahmed Monsur Borbhuiya , Madhava Yada Rao
IPC分类号： C07K16/00
CPC分类号： C07K14/61 , C07K1/20
摘要： Adding enough organic solvent to hydrophobic interactive chromatography elution buffer eliminates the need to also add detergent to the buffer when separating poly-peptides. For example, adding about 50% acetonitrile to detergent-free elution buffer enaibls one to separate full-length human growth hormone from its various truncated forms, to obtain hGH with a purity of >99.5%. This technique is useful to purify polypeptide where detergent to contamination in the resulting polypeptide is undesirable.
摘要翻译：向疏水交互色谱洗脱缓冲液中加入足够的有机溶剂不需要在分离多肽时也向缓冲液中加入洗涤剂。例如，将约50％的乙腈加入到不含洗涤剂的洗脱缓冲液中，从而将全长人生长激素与其各种截短形式分离，得到纯度> 99.5％的hGH。该技术可用于纯化多肽，其中所得多肽中的污染物的洗涤剂不期望。

9. 发明申请

US20110212508A1 Novel Synthetic Expression Vehicle 审中-公开
标题翻译：新型合成表达载体
公开(公告)号：US20110212508A1
公开(公告)日：2011-09-01
申请号：US13123960
申请日：2009-10-28
申请人： Laxmi Srinivas Rao , Shrikant Mishra , Milind Niphadkar , Priti Thakur , Genevieve Nazareth
发明人： Laxmi Srinivas Rao , Shrikant Mishra , Milind Niphadkar , Priti Thakur , Genevieve Nazareth
IPC分类号： C12N1/21 , C12N15/63 , C07H21/04
CPC分类号： C12N15/70
摘要： An expression vehicle comprising an isolated nucleic acid as shown in Seq ID No. 1 comprising of a synthetic hybrid promoter wherein the hybrid promoter comprises of an inducible arabinose promoter derived from E. coli and a synthetic stretch of 8-35 nucleotides from pHO regulon introduced in the region between −55 to −10 and ribosome binding site from the transcription initiation site.
摘要翻译：包含如Seq ID No.1所示的分离的核酸的表达载体，其包含合成的杂合启动子，其中所述杂合启动子包含来自大肠杆菌的诱导型阿拉伯糖启动子和来自pHO调节子的8-35个核苷酸的合成片段在-55至-10之间的区域和与转录起始位点的核糖体结合位点。

10. 发明申请

US20110189733A1 LOW CELL DENSITY FERMENTATION PROCESS FOR THE PRODUCTION OF HETEROLOGOUS RECOMBINANT PROTEINS IN MICROORGANISMS 审中-公开
标题翻译：用于生产微生物中异位重组蛋白的低密度发酵方法
公开(公告)号：US20110189733A1
公开(公告)日：2011-08-04
申请号：US13022617
申请日：2011-02-07
申请人： Milind Prabhakar NIPHADKAR , Genevieve Suzie Nazareth , Neelesh Ramesh Surlikar , Monsur Ahmed Borbhuiya , Uma Sankararaman , Dipanwita Maiti , Laxmi Srinivas Rao , Saptarshi Paul , Shrikant Mishra
发明人： Milind Prabhakar NIPHADKAR , Genevieve Suzie Nazareth , Neelesh Ramesh Surlikar , Monsur Ahmed Borbhuiya , Uma Sankararaman , Dipanwita Maiti , Laxmi Srinivas Rao , Saptarshi Paul , Shrikant Mishra
IPC分类号： C12P21/00 , C12N9/38
CPC分类号： C12P21/02 , C12N1/20
摘要： A low cell density fermentation process for the production of heterologous proteins in microorganisms. The cell culture obtained by cultivating host microorganisms transformed with a vector carrying genetic material for the said proteins and an inducible promoter under batch fermentation conditions is fed with a feed medium after an OD600 of 0.16 to 8 has been achieved or after 0 to 4 hrs from the start of the fermentation process. The feed medium comprises 5 to 30% of carbon source and 1 to 30% of nitrogen source and 0 mg to 400 mg antibiotics and 2.5 to 4.25% inorganic phosphates and trace elements. The concentration of the carbon source in the feed medium is 10 to 30 and the amino acid content in nitrogen source is 45 to 95%. The initial feed rate is in the range of 0 ml/hr to 12 ml/hr and is raised exponentially by an exponent in the range of 0.1 to 0.4 and/or linearly with the slope of the curve in the range of 0.5 to 3. The production of the heterologous proteins is induced with 0.01-4% inducer at a cell density of OD600 0.1-OD600 50. The feeding of the cell culture with the feed medium and the feed rate described above is continued after production has been induced. The pO2 is adjusted between 10% to 60% by passing sterile air into the fermentation broth and the temperature and pH of the fermentation broth are maintained at 33° C.-41° C. and 6.9-8.5, respectively during the entire fermentation.
摘要翻译：用于在微生物中产生异源蛋白质的低细胞密度发酵方法。在分批发酵条件下培养用携带所述蛋白质的遗传物质的载体转化的宿主微生物获得的细胞培养物在实现了OD600为0.16至8或0至4小时之后的饲料培养基中进料开始发酵过程。饲料培养基含有5-30％的碳源和1-10％的氮源和0mg-400mg的抗生素和2.5-4.25％的无机磷酸盐和微量元素。饲料培养基中碳源的浓度为10〜30，氮源中氨基酸含量为45〜95％。初始进料速率在0毫升/小时至12毫升/小时的范围内，并且在0.1至0.4范围内以指数指数增加和/或以0.5至3的范围内曲线的斜率线性地升高。以OD 600 0.1-OD 60050的细胞密度用0.01-4％诱导剂诱导异源蛋白的产生。在诱导生产后，继续用饲料培养基喂养细胞培养物和上述进料速率。通过将无菌空气送入发酵液中，将pO2调节在10％至60％之间，发酵液的温度和pH分别在整个发酵期间保持在33℃-41℃和6.9-8.5。

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