公开(公告)号：US20170183689A1

公开(公告)日：2017-06-29

申请号：US15224900

申请日：2016-08-01

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M.Y. Chen

IPC: C12P7/04 ,  C12P7/06 ,  C12N9/02

CPC classification number: C12P7/04 ,  C07K14/32 ,  C12N9/0042 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/06 ,  C12Y106/02004 ,  C12Y114/14001 ,  C12Y114/15 ,  C12Y114/15003 ,  Y02P20/52

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US10648006B2

公开(公告)日：2020-05-12

申请号：US15942001

申请日：2018-03-30

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M. Y. Chen

IPC: C12N9/02 ,  C12P7/04 ,  C07K14/32 ,  C12P7/06 ,  C12P7/02 ,  C12N15/53

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US09963720B2

公开(公告)日：2018-05-08

申请号：US15224900

申请日：2016-08-01

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M. Y. Chen

IPC: C12N9/02 ,  C12P7/04 ,  C12P7/06 ,  C12P7/02 ,  C12N15/53

CPC classification number: C12P7/04 ,  C07K14/32 ,  C12N9/0042 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/06 ,  C12Y106/02004 ,  C12Y114/14001 ,  C12Y114/15 ,  C12Y114/15003 ,  Y02P20/52

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US12258605B2

公开(公告)日：2025-03-25

申请号：US17560753

申请日：2021-12-23

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M. Y. Chen

IPC: C12N9/00 ,  C07K14/32 ,  C12N9/02 ,  C12P7/04 ,  C12P7/06

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US20220112524A1

公开(公告)日：2022-04-14

申请号：US17560753

申请日：2021-12-23

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M.Y. Chen

IPC: C12P7/04 ,  C07K14/32 ,  C12N9/02 ,  C12P7/06

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US09404096B2

公开(公告)日：2016-08-02

申请号：US14788365

申请日：2015-06-30

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M. Y. Chen

IPC: C12P7/04 ,  C12N9/02 ,  C07K14/32

CPC classification number: C12P7/04 ,  C07K14/32 ,  C12N9/0042 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/06 ,  C12Y106/02004 ,  C12Y114/14001 ,  C12Y114/15 ,  C12Y114/15003 ,  Y02P20/52

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US11214817B2

公开(公告)日：2022-01-04

申请号：US16872275

申请日：2020-05-11

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M. Y. Chen

IPC: C12N9/02 ,  C12N9/00 ,  C12N15/53 ,  C12P7/04 ,  C07K14/32 ,  C12P7/06

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US20180273983A1

公开(公告)日：2018-09-27

申请号：US15942001

申请日：2018-03-30

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M.Y. Chen

IPC: C12P7/04 ,  C12N9/02 ,  C12P7/06

CPC classification number: C12P7/04 ,  C07K14/32 ,  C12N9/0042 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/06 ,  C12Y106/02004 ,  C12Y114/14001 ,  C12Y114/15 ,  C12Y114/15003 ,  Y02E50/17 ,  Y02P20/52

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

公开(公告)号：US20150004668A1

公开(公告)日：2015-01-01

申请号：US14270268

申请日：2014-05-05

Applicant: The California Institute of Technology

Inventor： Frances Arnold ,  Peter Meinhold ,  Matthew W. Peters ,  Rudi Fasan ,  Mike M.Y. Chen

IPC: C12N9/02

CPC classification number: C12P7/04 ,  C07K14/32 ,  C12N9/0042 ,  C12N9/0071 ,  C12N9/0077 ,  C12P7/06 ,  C12Y106/02004 ,  C12Y114/14001 ,  C12Y114/15 ,  C12Y114/15003 ,  Y02P20/52

Abstract: This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

Abstract translation: 本发明涉及改性羟化酶。 本发明还涉及表达这种修饰的羟化酶的细胞和通过使合适的底物与这种细胞接触来制备羟基化烷烃的方法。

公开(公告)号：US20130273626A1

公开(公告)日：2013-10-17

申请号：US13673977

申请日：2012-11-09

Applicant: The California Institute of Technology

Inventor： Frances H. Arnold ,  Matthew W. Peters ,  Peter Meinhold

IPC: C12N9/00

CPC classification number: C12N9/0071 ,  C12N9/0077 ,  C12P7/04 ,  C12Y114/14001

Abstract: Cytochrome P450 BM-3 from Bacillus megaterium was engineered using a combination of directed evolution and site-directed mutagenesis to hydroxylate linear alkanes regio- and enantioselectively using atmospheric dioxygen as an oxidant. Mutant 9-10A-A328V hydroxylates octane primarily at the 2-positio to form S-2-octanol (40% ee). Another mutant, 1-12G, hydroxylates alkanes larger than hexane primarily at the 2-position, but forms R-2-alcohols (40-55% ee). These biocatalysts are highly active for alkane substrates and support thousands of product turnovers. These regio- and enantio-selectivities are retained in whole-cell biotransformations with E. coli, where the engineered P450s can be expressed at high levels and the expensive cofactor is supplied endogenously.

Abstract translation: 使用定向进化和定点诱变的组合将来自巨大芽孢杆菌的细胞色素P450 BM-3设计为使用大气二氧氧化物作为区域和对映选择性的羟基化线型烷烃。 突变体9-10A-A328V主要在2-位羟基化辛烷值以形成S-2-辛醇（40％ee）。 另一个突变体1-12G，主要在2-位羟基化烷烃大于己烷，但形成R-2-醇（40-55％ee）。 这些生物催化剂对烷烃底物具有高活性，并支持数千种产品周转。 这些区域和对映体选择性保留在与大肠杆菌的全细胞生物转化中，其中工程化的P450可以以高水平表达并且昂贵的辅因子是内源性提供的。