从嗜冷菌 Acinetobacter sp. MN 12 MTCC（10786）中提取和鉴定一种胞外低温活性及碱性稳定的蛋白酶。
Purification and Characterization of an Extracellular Low Temperature-Active and Alkaline Stable Peptidase from Psychrotrophic Acinetobacter sp. MN 12 MTCC (10786).
机构信息
CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176061 India.
出版信息
Indian J Microbiol. 2013 Mar;53(1):63-9. doi: 10.1007/s12088-012-0344-1. Epub 2012 Dec 30.
Abstract
An extracellular low temperature-active alkaline stable peptidase from Acinetobacter sp. MN 12 was purified to homogeneity with a purification fold of 9.8. The enzyme exhibited specific activity of 6,540 U/mg protein, with an apparent molecular weight of 35 kDa. The purified enzyme was active over broad range of temperature from 4 to 60 °C with optimum activity at 40 °C. The enzyme retained more than 75 % of activity over a broad range of pH (7.0-11.0) with optimum activity at pH 9.0. The purified peptidase was strongly inhibited by phenylmethylsulfonyl fluoride, giving an indication of serine type. The K m and V max for casein and gelatin were 0.3529, 2.03 mg/ml and 294.11, 384.61 μg/ml/min respectively. The peptidase was compatible with surfactants, oxidizing agents and commercial detergents, and effectively removed dried blood stains on cotton fabrics at low temperature ranging from 15 to 35 °C.
摘要
从不动杆菌 MN12 中分离得到一种胞外低温碱性稳定的肽酶，经纯化后其纯度达到 9.8 倍。该酶的比活为 6540 U/mg 蛋白，表观分子量为 35 kDa。纯化后的酶在 4 至 60°C 的宽温度范围内具有活性，最适温度为 40°C。该酶在 pH 7.0-11.0 的宽范围内保持超过 75%的活性，最适 pH 为 9.0。该肽酶被苯甲基磺酰氟强烈抑制，表明其属于丝氨酸类型。酪蛋白和明胶的 K m 和 V max 分别为 0.3529、2.03mg/ml 和 294.11、384.61μg/ml/min。该肽酶与表面活性剂、氧化剂和商业洗涤剂兼容，可有效去除棉织物上在 15 至 35°C 低温下的干涸血斑。
相似文献
1
Purification and Characterization of an Extracellular Low Temperature-Active and Alkaline Stable Peptidase from Psychrotrophic Acinetobacter sp. MN 12 MTCC (10786).
Indian J Microbiol. 2013 Mar;53(1):63-9. doi: 10.1007/s12088-012-0344-1. Epub 2012 Dec 30.
2
Purification and properties of detergent-compatible extracellular alkaline protease from Scopulariopsis spp.
Prep Biochem Biotechnol. 2014 Oct 3;44(7):738-59. doi: 10.1080/10826068.2013.854254.
3
Detergent-compatible, organic solvent-tolerant alkaline protease from Bacillus circulans MTCC 7942: Purification and characterization.
Prep Biochem Biotechnol. 2016;46(1):56-64. doi: 10.1080/10826068.2014.979205.
4
Detergent-, solvent- and salt-compatible thermoactive alkaline serine protease from halotolerant alkaliphilic Bacillus sp. NPST-AK15: purification and characterization.
Extremophiles. 2015 Sep;19(5):961-71. doi: 10.1007/s00792-015-0771-0. Epub 2015 Jul 10.
5
Molecular and biochemical characterization of an extracellular serine-protease from Vibrio metschnikovii J1.
J Ind Microbiol Biotechnol. 2009 Jul;36(7):939-48. doi: 10.1007/s10295-009-0572-5. Epub 2009 Apr 24.
6
Stability and detergent compatibility of a predominantly β-sheet serine protease from halotolerant B. aquimaris VITP4 strain.
Appl Biochem Biotechnol. 2014 Jan;172(2):687-700. doi: 10.1007/s12010-013-0524-4. Epub 2013 Oct 10.
7
Two step purification of Acinetobacter sp. lipase and its evaluation as a detergent additive at low temperatures.
Appl Biochem Biotechnol. 2008 Aug;150(2):139-56. doi: 10.1007/s12010-008-8143-1. Epub 2008 Feb 20.
8
Purification and characterization of alkaline soda-bleach stable protease from sp. APP-07 isolated from Laundromat soil.
J Genet Eng Biotechnol. 2018 Dec;16(2):273-279. doi: 10.1016/j.jgeb.2018.07.003. Epub 2018 Jul 20.
9
Purification and biochemical characterization of alkaline serine protease from Caesalpinia bonducella.
Nat Prod Commun. 2010 Jun;5(6):931-4.
10
Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6.
J Ind Microbiol Biotechnol. 2008 Feb;35(2):121-31. doi: 10.1007/s10295-007-0273-x. Epub 2007 Nov 10.
引用本文的文献
1
Biochemical features and biotechnological potential of a proteolytic extract from a psychrophilic Antarctic bacterium.
Braz J Microbiol. 2025 Jun;56(2):767-778. doi: 10.1007/s42770-024-01605-6. Epub 2025 Jan 29.
2
Cold-Active Enzymes and Their Potential Industrial Applications-A Review.
Molecules. 2022 Sep 10;27(18):5885. doi: 10.3390/molecules27185885.
3
Adaptation, production, and biotechnological potential of cold-adapted proteases from psychrophiles and psychrotrophs: recent overview.
J Genet Eng Biotechnol. 2020 Jul 28;18(1):36. doi: 10.1186/s43141-020-00053-7.
4
Bioprospecting Psychrotrophic Bacteria for Serine-Type Proteases from the Cold Areas of Western Himalayas.
Curr Microbiol. 2020 May;77(5):795-806. doi: 10.1007/s00284-020-01876-w. Epub 2020 Jan 10.
5
The Biodiversity of the Microbiota Producing Heat-Resistant Enzymes Responsible for Spoilage in Processed Bovine Milk and Dairy Products.
Front Microbiol. 2017 Mar 1;8:302. doi: 10.3389/fmicb.2017.00302. eCollection 2017.
6
Differential Response of Extracellular Proteases of Trichoderma Harzianum Against Fungal Phytopathogens.
Curr Microbiol. 2016 Sep;73(3):419-425. doi: 10.1007/s00284-016-1072-2. Epub 2016 Jun 8.
7
Draft Genome Sequence of Psychrotrophic Acinetobacter sp. Strain MN12 (MTCC 10786), Which Produces a Low-Temperature-Active and Alkaline-Stable Peptidase.
Genome Announc. 2014 Nov 20;2(6):e01167-14. doi: 10.1128/genomeA.01167-14.
8
Preliminary Extraction and Identification of the 44.5 kDa Outer Membrane Proteins Isolated from Bovine Fusobacterium necrophorum (AB).
Indian J Microbiol. 2013 Dec;53(4):395-9. doi: 10.1007/s12088-013-0388-x. Epub 2013 Mar 16.
本文引用的文献
1
Purification and characterization of a cold active alkaline protease from Stenotrophomonas sp., isolated from Kashmir, India.
World J Microbiol Biotechnol. 2012 Mar;28(3):1071-9. doi: 10.1007/s11274-011-0905-1. Epub 2011 Oct 1.
2
An on-demand metalloprotease from psychro-tolerant Exiguobacterium undae Su-1, the activity and stability of which are controlled by the Ca2+ concentration.
Biosci Biotechnol Biochem. 2012;76(5):986-92. doi: 10.1271/bbb.110997. Epub 2012 May 7.
3
Microbial proteases: detection, production, and genetic improvement.
Crit Rev Microbiol. 2011 Aug;37(3):262-76. doi: 10.3109/1040841X.2011.577029. Epub 2011 May 21.
4
Phylogenetic diversity of alkaline protease-producing psychrotrophic bacteria from glacier and cold environments of Lahaul and Spiti, India.
J Basic Microbiol. 2010 Apr;50(2):150-9. doi: 10.1002/jobm.200800400.
5
Proteases from psychrotrophs: an overview.
Crit Rev Microbiol. 2010 May;36(2):134-45. doi: 10.3109/10408410903485525.
6
A cold-active extracellular metalloprotease from Curtobacterium luteum (MTCC 7529): enzyme production and characterization.
J Gen Appl Microbiol. 2008 Dec;54(6):385-92. doi: 10.2323/jgam.54.385.
7
Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6.
J Ind Microbiol Biotechnol. 2008 Feb;35(2):121-31. doi: 10.1007/s10295-007-0273-x. Epub 2007 Nov 10.
8
Isolation of a psychrotrophic Exiguobacterium sp. SKPB5 (MTCC 7803) and characterization of its alkaline protease.
Curr Microbiol. 2007 Mar;54(3):224-9. doi: 10.1007/s00284-006-0402-1.
9
Purification and characterization of an extracellular cold-active serine protease from the psychrophilic bacterium Colwellia sp. NJ341.
Biotechnol Lett. 2005 Aug;27(16):1195-8. doi: 10.1007/s10529-005-0016-x.
10
Microbial proteases in peptide synthesis: approaches and applications.
Appl Microbiol Biotechnol. 2005 Oct;68(6):726-36. doi: 10.1007/s00253-005-0094-7. Epub 2005 Oct 13.
Zotero 插件