公开(公告)号：US6077955A

公开(公告)日：2000-06-20

申请号：US108728

申请日：1998-07-01

Applicant: Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

Inventor： Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

IPC: C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D211/76 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/00 ,  C12P17/10 ,  C07D211/60 ,  C07D207/00

CPC classification number: C07D211/76 ,  C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/002 ,  C12P13/02 ,  C12P17/10 ,  C12R1/01 ,  Y02P20/52

Abstract: A process for the preparation of five-membered or six-membered ring lactams from aliphatic .alpha.,.omega.-dinitriles has been developed. In the process an aliphatic .alpha.,.omega.-dinitrile is first converted to an ammonium salt of an .omega.-nitrilecarboxylic acid in aqueous solution using a catalyst having an aliphatic nitrilase (EC 3.5.5.7) activity, or a combination of nitrile hydratase (EC 4.2.1.84) and amidase (EC 3.5.1.4) activities. The ammonium salt of the .omega.-nitrilecarboxylic acid is then converted directly to the corresponding lactam by hydrogenation in aqueous solution, without isolation of the intermediate .omega.-nitrilecarboxylic acid or .omega.-aminocarboxylic acid. When the aliphatic .alpha.,.omega.-dinitrile is also unsymmetrically substituted at the .alpha.-carbon atom, the nitrilase produces the .omega.-nitrilecarboxylic acid ammonium salt resulting from hydrolysis of the .omega.-nitrile group with greater than 98% regioselectivity, thereby producing only one of the two possible lactam products during the subsequent hydrogenation. A heat-treatment process to select for desirable regioselective nitrilase or nitrile hydratase activities while destroying undesirable activities is also provided.

Abstract translation: 已经开发了从脂肪族α，ω-二腈制备五元或六元环内酰胺的方法。 在该方法中，使用具有脂族腈水解酶（EC 3.5.5.7）活性的催化剂或腈水合酶（EC 4.2）的组合，首先将脂族α，ω-二腈转化为ω-二硝基甲酸的铵盐 .1.84）和酰胺酶（EC 3.5.1.4）活性。 然后通过在水溶液中氢化将ω-二硝基羧酸的铵盐直接转化成相应的内酰胺，而不分离中间体ω-二硝基甲酸或ω-氨基羧酸。 当脂肪族α，ω-二腈在α-碳原子处也不对称取代时，腈水解酶产生由ω-腈基水解产生的ω-亚硝基羧酸铵盐，其具有大于98％的区域选择性，从而仅产生 在随后氢化期间两种可能的内酰胺产物。 还提供了在破坏不期望的活性的同时选择期望的区域选择性腈水解酶或腈水合酶活性的热处理方法。

公开(公告)号：US5908954A

公开(公告)日：1999-06-01

申请号：US108445

申请日：1998-07-01

Applicant: Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

Inventor： Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

IPC: C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D211/76 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/00 ,  C12P17/10 ,  C07C255/03 ,  C12P17/12

CPC classification number: C07D211/76 ,  C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/002 ,  C12P13/02 ,  C12P17/10 ,  C12R1/01 ,  Y02P20/52

Abstract: A process for the preparation of five-membered or six-membered ring lactams from aliphatic .alpha.,.omega.-dinitriles has been developed. In the process an aliphatic .alpha.,.omega.-dinitrile is first converted to an ammonium salt of an .omega.-nitrile-carboxylic acid in aqueous solution using a catalyst having an aliphatic nitrilase (EC 3.5.5.7) activity, or a combination of nitrile hydratase (EC 4.2.1.84) and arnidase (EC 3.5.1.4) activities. The ammonium salt of the .omega.-nitrilecarboxylic acid is then converted directly to the corresponding lactam by hydrogenation in aqueous solution, without isolation of the intermediate .omega.-nitrilecarboxylic acid or .omega.-aminocarboxylic acid. When the aliphatic .alpha.,.omega.-dinitrile is also unsymmetrically substituted at the .alpha.-carbon atom, the nitrilase produces the .omega.-nitrilecarboxylic acid ammonium salt resulting from hydrolysis of the .omega.-nitrile group with greater than 98% regioselectivity, thereby producing only one of the two possible lactam products during the subsequent hydrogenation. A heat-treatment process to select for desirable regioselective nitrilase or nitrile hydratase activities while destroying undesirable activities is also provided.

Abstract translation: 已经开发了从脂肪族α，ω-二腈制备五元或六元环内酰胺的方法。 在该方法中，首先使用具有脂族腈水解酶（EC 3.5.5.7）活性的催化剂或腈水合酶的组合（在水溶液中将ω-腈 - 羧酸的铵盐转化为ω-腈 - 羧酸的铵盐， EC 4.2.1.84）和arnidase（EC 3.5.1.4）活动。 然后通过在水溶液中氢化将ω-二硝基羧酸的铵盐直接转化成相应的内酰胺，而不分离中间体ω-二硝基甲酸或ω-氨基羧酸。 当脂肪族α，ω-二腈在α-碳原子处也不对称取代时，腈水解酶产生由ω-腈基水解产生的ω-亚硝基羧酸铵盐，其具有大于98％的区域选择性，从而仅产生 在随后氢化期间两种可能的内酰胺产物。 还提供了在破坏不期望的活性的同时选择期望的区域选择性腈水解酶或腈水合酶活性的热处理方法。

公开(公告)号：US5656489A

公开(公告)日：1997-08-12

申请号：US592243

申请日：1996-01-26

Applicant: Robert Donald Fallon

Inventor： Robert Donald Fallon

IPC: A62D3/02 ,  A62D3/00 ,  B09C1/10 ,  C02F3/28

CPC classification number: B09C1/10 ,  A62D3/02 ,  A62D2101/24

Abstract: In contaminated media, transformation of organoleads, especially tetraethyllead, into inorganic form is achieved by stimulation of indigenous microbial populations. Complex biological extracts and other nutrient amendments promote microbial activity under aerobic or anaerobic conditions. Under anaerobic conditions, sulfide production by sulfate reducing bacteria is especially promoted. Transformation to the inorganic form reduces lead toxicity and mobility in ground water supplies.

Abstract translation: 在受污染的媒介中，通过刺激本地微生物群落实现了有机物，特别是四乙基铅的转化为无机形式。 复杂的生物提取物和其他营养物质的修正促进有氧或厌氧条件下的微生物活动。 在厌氧条件下，特别促进硫酸盐还原菌的硫化物生成。 转化为无机形式降低了地下水供应中的铅毒性和流动性。

公开(公告)号：US06251650B1

公开(公告)日：2001-06-26

申请号：US09687594

申请日：2000-10-13

Applicant: Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

Inventor： Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

IPC: C12N978

CPC classification number: C12P13/02 ,  C12N9/78 ,  C12N9/80 ,  C12P7/40 ,  C12P17/00 ,  C12P41/006 ,  Y10S435/877

Abstract: The present invention provides a nitrile hydratase nucleic acid fragment isolated from Pseudomonas putida which encodes a nitrile hydratase activity capable of catalyzing the hydrolysis of certain racemic nitriles to the corresponding R- or S-amides. Also provided are transformed microorganisms capable of the active expression of said nitrile hydratase activity. Additionally, the invention provides a transformant harboring the nitrile hydratase gene in conjunction with an amidase gene, both of which may be co-expressed producing active nitrile hydratase and amidase enzymes respectively. Methods for the production of such enantiomeric materials are also provided.

Abstract translation: 本发明提供从恶臭假单胞菌（Pseudomonas putida）分离的腈水合酶核酸片段，其编码能够将某些外消旋腈水解成相应的R-或S-酰胺的腈水合酶活性。 还提供了能够主动表达所述腈水合酶活性的转化微生物。 另外，本发明提供了一种携带腈水合酶基因的转化体，其与酰胺酶基因结合，两者都可以分别共同表达产生活性腈水合酶和酰胺酶。 还提供了生产这种对映体材料的方法。

公开(公告)号：US6066490A

公开(公告)日：2000-05-23

申请号：US987938

申请日：1997-12-10

Applicant: Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

Inventor： Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

IPC: C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D211/76 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/00 ,  C12P17/10 ,  C12N1/12

CPC classification number: C07D211/76 ,  C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/002 ,  C12P13/02 ,  C12P17/10 ,  C12R1/01 ,  Y02P20/52

Abstract: A process for the preparation of five-membered or six-membered ring lactams from aliphatic .alpha.,.omega.-dinitriles has been developed. In the process an aliphatic .alpha.,.omega.-dinitrile is first converted to an ammonium salt of an .omega.-nitrilecarboxylic acid in aqueous solution using a catalyst having an aliphatic nitrilase (EC 3.5.5.7) activity, or a combination of nitrile hydratase (EC 4.2.1.84) and amidase (EC 3.5.1.4) activities. The ammonium salt of the .omega.-nitrilecarboxylic acid is then converted directly to the corresponding lactam by hydrogenation in aqueous solution, without isolation of the intermediate .omega.-nitrilecarboxylic acid or .omega.-aminocarboxylic acid. When the aliphatic .alpha.,.omega.-dinitrile is also unsymmetrically substituted at the .alpha.-carbon atom, the nitrilase produces the .omega.-nitrilecarboxylic acid ammonium salt resulting from hydrolysis of the .omega.-nitrile group with greater than 98% regioselectivity, thereby producing only one of the two possible lactam products during the subsequent hydrogenation. A heat-treatment process to select for desirable regioselective nitrilase or nitrile hydratase activities while destroying undesirable activities is also provided.

Abstract translation: 已经开发了从脂肪族α，ω-二腈制备五元或六元环内酰胺的方法。 在该方法中，使用具有脂族腈水解酶（EC 3.5.5.7）活性的催化剂或腈水合酶（EC 4.2）的组合，首先将脂族α，ω-二腈转化为ω-二硝基甲酸的铵盐 .1.84）和酰胺酶（EC 3.5.1.4）活性。 然后通过在水溶液中氢化将ω-二硝基羧酸的铵盐直接转化成相应的内酰胺，而不分离中间体ω-二硝基甲酸或ω-氨基羧酸。 当脂肪族α，ω-二腈在α-碳原子处也不对称取代时，腈水解酶产生由ω-腈基水解产生的ω-亚硝基羧酸铵盐，其具有大于98％的区域选择性，从而仅产生 在随后氢化期间两种可能的内酰胺产物。 还提供了在破坏不期望的活性的同时选择期望的区域选择性腈水解酶或腈水合酶活性的热处理方法。

公开(公告)号：US5936114A

公开(公告)日：1999-08-10

申请号：US108631

申请日：1998-07-01

Applicant: Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

Inventor： Robert Di Cosimo ,  Robert Donald Fallon ,  John Edward Gavagan ,  Frank Edward Herkes

IPC: C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D211/76 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/00 ,  C12P17/10 ,  C07C255/29

CPC classification number: C07D211/76 ,  C07C255/19 ,  C07C255/23 ,  C07D201/08 ,  C07D207/26 ,  C07D223/10 ,  C12N9/78 ,  C12N9/88 ,  C12P13/002 ,  C12P13/02 ,  C12P17/10 ,  C12R1/01 ,  Y02P20/52

Abstract: A process for the preparation of five-membered or six-membered ring lactams from aliphatic .alpha.,.omega.-dinitriles has been developed. In the process an aliphatic .alpha.,.omega.-dinitrile is first converted to an ammonium salt of an .omega.-nitrilecarboxylic acid in aqueous solution using a catalyst having an aliphatic nitrilase (EC 3.5.5.7) activity, or a combination of nitrile hydratase (EC 4.2.1.84) and amidase (EC 3.5.1.4) activities. The ammonium salt of the .omega.-nitrilecarboxylic acid is then converted directly to the corresponding lactam by hydrogenation in aqueous solution, without isolation of the intermediate .omega.-nitrilecarboxylic acid or .omega.-aminocarboxylic acid. When the aliphatic .alpha.,.omega.-dinitrile is also unsymmetrically substituted at the .alpha.-carbon atom, the nitrilase produces the .omega.-nitrilecarboxylic acid ammonium salt resulting from hydrolysis of the .omega.-nitrile group with greater than 98% regioselectivity, thereby producing only one of the two possible lactam products during the subsequent hydrogenation. A heat-treatment process to select for desirable regioselective nitrilase or nitrile hydratase activities while destroying undesirable activities is also provided.

Abstract translation: 已经开发了从脂肪族α，ω-二腈制备五元或六元环内酰胺的方法。 在该方法中，使用具有脂族腈水解酶（EC 3.5.5.7）活性的催化剂或腈水合酶（EC 4.2）的组合，首先将脂族α，ω-二腈转化为ω-二硝基甲酸的铵盐 .1.84）和酰胺酶（EC 3.5.1.4）活性。 然后通过在水溶液中氢化将ω-二硝基羧酸的铵盐直接转化成相应的内酰胺，而不分离中间体ω-二硝基甲酸或ω-氨基羧酸。 当脂肪族α，ω-二腈在α-碳原子处也不对称取代时，腈水解酶产生由ω-腈基水解产生的ω-亚硝基羧酸铵盐，其具有大于98％的区域选择性，从而仅产生 在随后氢化期间两种可能的内酰胺产物。 还提供了在破坏不期望的活性的同时选择期望的区域选择性腈水解酶或腈水合酶活性的热处理方法。

公开(公告)号：US06368804B1

公开(公告)日：2002-04-09

申请号：US09614914

申请日：2000-07-12

Applicant: Arie Ben-Bassat ,  Robert Dicosimo ,  Robert Donald Fallon

Inventor： Arie Ben-Bassat ,  Robert Dicosimo ,  Robert Donald Fallon

IPC: C12N120

CPC classification number: C12N11/10 ,  C12N1/04 ,  C12N9/78 ,  C12N9/96 ,  C12N11/04 ,  C12N11/08 ,  C12P13/00

Abstract: A method for preserving immobilized or unimmobilized microbial cells having nitrilase activity and for stabilizing the nitrilase activity of unimmobilized or immobilized microbial cells has been developed. The unimmobilized or immobilized microbial cells are stored in an aqueous solution containing from about 0.10 M to the saturation concentration of an inorganic salt of bicarbonate or carbonate, including ammonium, sodium and potassium salts of bicarbonate or carbonate. Aqueous suspensions containing at least 100 mM bicarbonate or carbonate limit microbial contamination of the stored enzyme catalyst, as well as stabilize the desired nitrilase activity of the uninmmobilized or immobilized cells. Microorganisms which are characterized by an nitrilase activity and are stabilized and preserved by this method include Acidovorax facilis 72-PF-15 (ATCC 55747), Acidovorax facilis 72-PF-17 (ATCC 55745), Acidovorax facilis 72W (ATCC 55746), and transformed microbial cells having nitrilase activity, preferably E. coli SS1001 (ATCC PTA-1177) which is transformed with Acidovorax facilis 72W nitrilase activity.

Abstract translation: 已经开发了用于保存具有腈水解酶活性的固定或未固化的微生物细胞并稳定未固定或固定的微生物细胞的腈水解酶活性的方法。 将未固定或固定的微生物细胞储存在含有约0.10M的水溶液中，使饱和浓度为碳酸氢盐或碳酸盐的无机盐，包括碳酸氢盐或碳酸盐的铵，钠和钾盐。 含有至少100mM碳酸氢盐或碳酸氢盐的水性悬浮液限制所储存的酶催化剂的微生物污染物，以及稳定未固定化或固定的细胞的所需腈水解酶活性。 其特征在于腈水解酶活性并通过该方法稳定和保存的微生物包括Acidovorax facilis 72-PF-15（ATCC 55747），Acidovorax facilis 72-PF-17（ATCC 55745），Acidovorax facilis 72W（ATCC 55746）和 转化的具有腈水解酶活性的微生物细胞，优选用Acidovorax facilis 72W腈水解酶活性转化的大肠杆菌SS1001（ATCC PTA-1177）。

公开(公告)号：US5888785A

公开(公告)日：1999-03-30

申请号：US103411

申请日：1998-06-24

Applicant: Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

Inventor： Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

IPC: C12N15/09 ,  C07K14/21 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N9/78 ,  C12N9/80 ,  C12N15/00 ,  C12N15/55 ,  C12P7/40 ,  C12P13/02 ,  C12P17/00 ,  C12P41/00 ,  C12R1/19 ,  C12R1/40 ,  C12R1/84 ,  C12N9/88

CPC classification number: C12P13/02 ,  C12N9/78 ,  C12N9/80 ,  C12P17/00 ,  C12P41/006 ,  C12P7/40 ,  Y10S435/877

Abstract: The present invention provides a nitrile hydratase nucleic acid fragment isolated from Pseudomonas putida which encodes a nitrile hydratase activity capable of catalyzing the hydrolysis of certain racemic nitrites to the corresponding R- or S-amides. Also provided are transformed microorganisms capable of the active expression of said nitrile hydratase activity. Additionally, the invention provides a transformant harboring the nitrile hydratase gene in conjunction with an amidase gene, both of which may be co-expressed producing active nitrile hydratase and amidase enzymes respectively. Methods for the production of such enantiomeric materials are also provided.

Abstract translation: 本发明提供从恶臭假单胞菌（Pseudomonas putida）分离的腈水合酶核酸片段，其编码能够将某些外消旋亚硝酸盐水解成相应的R-或S-酰胺的腈水合酶活性。 还提供了能够主动表达所述腈水合酶活性的转化微生物。 另外，本发明提供了一种携带腈水合酶基因的转化体，其与酰胺酶基因结合，两者都可以分别共同表达产生活性腈水合酶和酰胺酶。 还提供了生产这种对映体材料的方法。

公开(公告)号：US5811286A

公开(公告)日：1998-09-22

申请号：US726136

申请日：1996-10-04

Applicant: Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

Inventor： Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

IPC: C12N15/09 ,  C07K14/21 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N9/78 ,  C12N9/80 ,  C12N15/00 ,  C12N15/55 ,  C12P7/40 ,  C12P13/02 ,  C12P17/00 ,  C12P41/00 ,  C12R1/19 ,  C12R1/40 ,  C12R1/84 ,  C12N1/20 ,  C07H21/04 ,  C12N9/88

CPC classification number: C12P13/02 ,  C12N9/78 ,  C12N9/80 ,  C12P17/00 ,  C12P41/006 ,  C12P7/40 ,  Y10S435/877

Abstract: The present invention provides a nitrile hydratase nucleic acid fragment isolated from Pseudomonas putida which encodes a nitrile hydratase activity capable of catalyzing the hydrolysis of certain racemic nitriles to the corresponding R- or S-amides. Also provided are transformed microorganisms capable of the active expression of said nitrile hydratase activity. Additionally, the invention provides a transformant harboring the nitrile hydratase gene in conjunction with an amidase gene, both of which may be co-expressed producing active nitrile hydratase and amidase enzymes respectively. Methods for the production of such enantiomeric materials are also provided.

Abstract translation: 本发明提供从恶臭假单胞菌（Pseudomonas putida）分离的腈水合酶核酸片段，其编码能够将某些外消旋腈水解成相应的R-或S-酰胺的腈水合酶活性。 还提供了能够主动表达所述腈水合酶活性的转化微生物。 另外，本发明提供了一种携带腈水合酶基因的转化体，其与酰胺酶基因结合，两者都可以分别共同表达产生活性腈水合酶和酰胺酶。 还提供了生产这种对映体材料的方法。

公开(公告)号：US6133421A

公开(公告)日：2000-10-17

申请号：US103434

申请日：1998-06-24

Applicant: Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

Inventor： Robert Donald Fallon ,  Mark James Nelson ,  Mark Scott Payne

IPC: C12N15/09 ,  C07K14/21 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N9/78 ,  C12N9/80 ,  C12N15/00 ,  C12N15/55 ,  C12P7/40 ,  C12P13/02 ,  C12P17/00 ,  C12P41/00 ,  C12R1/19 ,  C12R1/40 ,  C12R1/84 ,  C07H21/04 ,  C12N1/00 ,  C12N1/20 ,  C12N9/88

CPC classification number: C12P13/02 ,  C12N9/78 ,  C12N9/80 ,  C12P17/00 ,  C12P41/006 ,  C12P7/40 ,  Y10S435/877

Abstract: 350 The present invention provides a nitrile hydratase nucleic acid fragment isolated from Pseudomonas putida which encodes a nitrile hydratase activity capable of catalyzing the hydrolysis of certain racemic nitriles to the corresponding R- or S-amides. Also provided are transformed microorganisms capable of the active expression of said nitrile hydratase activity. Additionally, the invention provides a transformant harboring the nitrile hydratase gene in conjunction with an amidase gene, both of which may be co-expressed producing active nitrile hydratase and amidase enzymes respectively. Methods for the production of such enantiomeric materials are also provided.

Abstract translation: 350本发明提供从恶臭假单胞菌分离的腈水合酶核酸片段，其编码能够催化某些外消旋腈水解成相应的R-或S-酰胺的腈水合酶活性。 还提供了能够主动表达所述腈水合酶活性的转化微生物。 另外，本发明提供了一种携带腈水合酶基因的转化体，其与酰胺酶基因结合，两者都可以分别共同表达产生活性腈水合酶和酰胺酶。 还提供了生产这种对映体材料的方法。