A propionitrile-induced nitrilase of Rhodococcus sp NDB 1165 and its application in nicotinic acid synthesis

被引:45
作者
Prasad, Shreenath
Misra, Anurag
Jangir, Ved Prakash
Awasthi, Abhishek
Raj, Jog
Bhalla, Tek Chand
机构
[1] Himachal Pradesh Univ, Dept Biotechnol, Shimla 171005, Himachal Prades, India
[2] Seedling Acad Design Technol & Management, Jaipur 302025, Rajasthan, India
[3] Saroj Inst Technol & Management, Lucknow 226010, Uttar Pradesh, India
关键词
3-cyanopyridine; fed-batch reaction; nicotinic acid; nitrilase; Rhodococcus sp; NDB; 1165;
D O I
10.1007/s11274-006-9230-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Rhodococcus sp. NDB 1165, a nitrile-transforming organism was isolated from temperate forest soil of Himalayas. The nitrilase (EC 3.5.5.2) activity of this organism had higher substrate specificity toward aromatic nitriles (benzonitrile, 3-cyanopyridine and 4-cyanopyridine) and unsaturated aliphatic nitrile (acrylonitrile) in comparison to saturated aliphatic nitriles (acetonitrile, propionitrile, butyronitrile and isobutyronitrile) nitrile and arylacetonitrile (phenylacetonitrile and indole-3-acetonitrile). The nitrilase of Rhodococcus sp. NDB 1165 was inducible in nature and propionitrile proved to be an efficient inducer. However, the salts of ferrous and cobalt ions had an inhibitory effect. Under optimized reaction conditions (pH 8.0 and temperature 45 degrees C) the nitrilase activity of this organism was 2.39 +/- 0.07 U/mg dry cell mass (dcm). The half-life of this enzyme was 150 min and 40 min at 45 degrees C and 50 degrees C respectively. However, it was quite stable at 40 degrees C and around 58 % activity was retained even after 6 h at this temperature. The V (max) and K (m) value of this nitrilase were 1.67 mu mol/ml min and 0.1 M respectively using 3-cyanopyridine as substrate. However, the decrease in V (max) and K (m) values (0.56 mu mol/ml min and 0.02 M, respectively) were observed at > 0.05 M 3-cyanopyridine which revealed that this enzyme experienced uncompetitive inhibition at higher substrate concentrations. Under optimized reaction conditions, 1.6 M 3-cyanopyridine was successfully converted in to nicotinic acid using 2.0 mg resting cells (dcm)/ml reaction mixture in 11 h. This is the highest production of nicotinic acid i.e. 8.95 mg/mg resting cells (dcm)/h as compared to nitrilase systems reported hitherto.
引用
收藏
页码:345 / 353
页数:9
相关论文
共 28 条
[1]   Thermostable nitrilase catalysed production of nicotinic acid from 3-cyanopyridine [J].
Almatawah, QA ;
Cowan, DA .
ENZYME AND MICROBIAL TECHNOLOGY, 1999, 25 (8-9) :718-724
[2]   Characterization of an inducible nitrilase from a thermophilic bacillus [J].
Almatawah, QA ;
Cramp, R ;
Cowan, DA .
EXTREMOPHILES, 1999, 3 (04) :283-291
[3]   RETRACTED: The nitrile-degrading enzymes: current status and future prospects (Retracted article. See vol. 100, pg. 7359, 2016) [J].
Banerjee, A ;
Sharma, R ;
Banerjee, UC .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2002, 60 (1-2) :33-44
[4]  
BELITZ HD, 1999, FOOD CHEM, P423
[5]  
Bhalla TC, 2005, CAN J MICROBIOL, V51, P705, DOI [10.1139/w05-046, 10.1139/W05-046]
[6]  
BHALLA TC, 1992, APPL MICROBIOL BIOT, V37, P184
[7]  
BLAKEY AJ, 1995, FEMS MICROBIOL LETT, V129, P57
[8]  
BOLLAG DM, 1991, PROTEIN METHODS, P1
[9]  
Chaplin MF, 1990, ENZYME TECHNOLOGY, P1
[10]   Biochemistry and biotechnology of mesophilic and thermophilic nitrile metabolizing enzymes [J].
Cowan, D ;
Cramp, R ;
Pereira, R ;
Graham, D ;
Almatawah, Q .
EXTREMOPHILES, 1998, 2 (03) :207-216