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    RECOMBINANT CELLS PRODUCING ITACONIC ACID AND METHYL ESTERS THEREOF
    12.
    发明申请
    RECOMBINANT CELLS PRODUCING ITACONIC ACID AND METHYL ESTERS THEREOF 审中-公开

    公开(公告)号：US20170191091A1

    公开(公告)日：2017-07-06

    申请号：US15314494

    申请日：2015-05-28

    Applicant: DSM IP Assets B.V.

    Inventor： Zheng ZHAO ,  Bernard MEIJRINK ,  Robertus Antonius Mijndert VAN DER HOEVEN ,  Liang WU ,  Johannes Andries ROUBOS ,  Marina VAN GELDER

    IPC: C12P7/62 ,  C12N15/52 ,  C12P7/46

    CPC classification number: C12P7/62 ,  C12N15/52 ,  C12P7/46

    Abstract: The present invention relates to a recombinant cell which is capable of producing one or more of itaconic acid, 4-methyl itaconate or 1-methyl itaconate, wherein the said recombinant microorganism has been modified in its genome such that it results in a deficiency in the production of a trans-aconitate methyltransferase. A recombinant yeast cell which is capable of producing itaconic acid and which overexpresses: —a nucleic acid encoding a polypeptide having cis-aconitate decarboxylase activity; and —one or more nucleic acids encoding polypeptides which separately or together catalyze a reaction towards acetyl CoA, wherein the said recombinant microorganism has been modified in its genome such that it results in a deficiency in the production of a trans-aconitate methyltransferase. These recombinant yeast cells may be used in processes for the production of itaconic acid, 4-methyl itaconate or 1-methyl itaconate.

    GUIDE RNA ASSEMBLY VECTOR
    16.
    发明申请
    GUIDE RNA ASSEMBLY VECTOR 审中-公开

    公开(公告)号：US20190055544A1

    公开(公告)日：2019-02-21

    申请号：US15741985

    申请日：2016-07-06

    Applicant: DSM IP ASSETS B.V.

    Inventor： Paulus Petrus DE WAAL ,  Johannes Andries ROUBOS ,  Paul KLAASSEN ,  René VERWAAL

    IPC: C12N15/10 ,  C12N15/62 ,  C12N15/81 ,  C12N15/64 ,  C12Q1/6806 ,  C12Q1/686

    Abstract: The present invention relates relates to a CRISPR-CAS system for a host cell, in particular to a method to produce a circular vector comprising one or more guide-polynucleotide expression cassettes, wherein said one or more guide-polynucleotide expression cassettes comprise a polynucleotide encoding a guide-polynucleotide operably linked to one or more control sequences which direct the expression of said guide-polynucleotide in a host cell, wherein said guide-polynucleotide comprises a guide-sequence that essentially is the reverse complement of a target-polynucleotide in a host cell and wherein the guide-polynucleotide can direct binding of a Cas protein at a target-polynucleotide in a host cell to form a CRISPR-Cas complex, in vivo.

    Method for Achieving Improved Polypeptide Expression
    18.
    发明申请
    Method for Achieving Improved Polypeptide Expression 审中-公开
    实现改良多肽表达的方法

    公开(公告)号：US20140377800A1

    公开(公告)日：2014-12-25

    申请号：US14281934

    申请日：2014-05-20

    Applicant: DSM IP ASSETS B.V.

    Inventor： Johannes Andries ROUBOS ,  Noel Nicolaas Maria Elisabeth VAN PEIJ

    IPC: C12N15/67 ,  C12N9/92 ,  C12N9/30

    CPC classification number: C12N15/67 ,  C12N9/242 ,  C12N9/92 ,  C12N15/80 ,  C12N15/81 ,  C12N2800/22

    Abstract: The present invention relates to methods of optimization of a protein coding sequences for expression in a given host cell. The methods apply genetic algorithms to optimise single codon fitness and/or codon pair fitness sequences coding for a predetermined amino acid sequence. In the algorithm generation of new sequence variants and subsequent selection of fitter variants is reiterated until the variant coding sequences reach a minimum value for single codon fitness and/or codon pair fitness. The invention also relates to a computer comprising a processor and memory, the processor being arranged to read from and write into the memory, the memory comprising data and instructions arranged to provide the processor with the capacity to perform the genetic algorithms for optimisation of single codon fitness and/or codon pair fitness. The invention further relates to nucleic acids comprising a coding sequence for a predetermined amino acid sequence, the coding sequence being optimised with respect to single codon fitness and/or codon pair fitness for a given host in the methods of the invention, to host cells comprising such nucleic acids and to methods for producing polypeptides and other fermentation products in which these host cells are used.

    Abstract translation: 本发明涉及用于在给定宿主细胞中表达的蛋白质编码序列的优化方法。 该方法应用遗传算法来优化编码预定氨基酸序列的单个密码子适合度和/或密码子对适应性序列。 在算法中重复生成新的序列变体并随后选择拟合变体，直到变体编码序列达到单个密码子适应度和/或密码子对适应度的最小值。 本发明还涉及一种包括处理器和存储器的计算机，所述处理器被布置为从存储器读取和写入存储器，所述存储器包括数据和指令，所述数据和指令被布置为向处理器提供执行用于优化单个密码子的遗传算法的能力 健身和/或密码子对健身。 本发明还涉及包含用于预定氨基酸序列的编码序列的核酸，所述编码序列针对本发明方法中给定宿主的单个密码子适合度和/或密码子对适应性优化至包含 这种核酸以及用于产生其中使用这些宿主细胞的多肽和其它发酵产物的方法。

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