Screening and characterization of proteases produced by deep-sea cold seep bacteria

Chenchen GUO; Chaomin SUN; Shimei WU

doi:10.1007/s00343-021-0441-2

Your Location：

Home >

Browse articles >

Screening and characterization of proteases produced by deep-sea cold seep bacteria

Biology | Updated：2023-05-20

- Screening and characterization of proteases produced by deep-sea cold seep bacteria
- Screening and characterization of proteases produced by deep-sea cold seep bacteria
- 海洋湖沼学报（英文） 2022年40卷第2期页码：678-689
- Affiliations：
  
  1.College of Life Sciences, Qingdao University, Qingdao 266071, China
  2.CAS Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
- Author bio：
  
  Shimei WU, E-mail: shimeiwu2016@126.com
- Funds：
  
  the National Natural Science Foundation of China(31600035);the China Ocean Mineral Resources R & D Association Grant(DY135-B2-14);© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2022
- DOI：10.1007/s00343-021-0441-2
  中图分类号：
- 收稿：2020-11-20，
  
  录用：2021-2-27，
  
  网络首发：2021-04-30，
  
  纸质出版：2022-03
- Accepted：
Scan QR Code
Screening and characterization of proteases produced by deep-sea cold seep bacteria[J]. 海洋湖沼学报（英文）, 2022,40(2):678-689.

Chenchen GUO, Chaomin SUN, Shimei WU. Screening and characterization of proteases produced by deep-sea cold seep bacteria[J]. Journal of Oceanology and Limnology, 2022, 40(2): 678-689.
Screening and characterization of proteases produced by deep-sea cold seep bacteria[J]. 海洋湖沼学报（英文）, 2022,40(2):678-689. DOI： 10.1007/s00343-021-0441-2.

Chenchen GUO, Chaomin SUN, Shimei WU. Screening and characterization of proteases produced by deep-sea cold seep bacteria[J]. Journal of Oceanology and Limnology, 2022, 40(2): 678-689. DOI： 10.1007/s00343-021-0441-2.

摘要

Abstract

Fifty protease-producing strains were screened from sediment of deep-sea cold seep

and divided into four different categories:

Bacillus

Pseudoalteromonas

Vibrio

and

Alteromonas

according to the sequences of 16s rRNA. Their abilities to produce protease

amylase

and lipase were determined

and a

Bacillus

strain gcc-1 displayed very strong alkaline protease activity and stability under different thermal and acidic conditions. The purification of the protease produced by strain gcc-1 was carried out by precipitation with ammonium sulfate

and sequentially chromatographed by anion exchange column and gel filtration. The purified protease showed a single band at the molecular weight of 28 kDa by SDS-PAGE. The characterization results show that the purified protease exhibited a considerable activity and stability in a wide thermal range of 10-80℃ and a wide acidic range of pH 6.5-11.5

and displayed highest activity at 40℃ and pH 8.5. Notably

the protease still maintained high activity even at low to 10℃. Furthermore

the protease exhibited good stability in presence of different surfactants

organic solvents

oxidizing agent H

and commercial detergents. Therefore

the protease produced by gcc-1 is a cold active and high stable enzyme

and has a promising potential in laundry detergent as an additive.

关键词

Keywords

references

Ali M B, Khemakhem B, Robert X, Haser R, Bejar S. 2006. Thermostability enhancement and change in starch hydrolysis profile of the maltohexaose-forming amylase of Bacillus stearothermophilus US100 strain. Biochemical Journal , 394 (1): 51–56, https://doi.org/10.1042/BJ20050726..

Cristobal H A, López M A, Kothe E, Abate C M. 2011. Diversity of protease-producing marine bacteria from sub-Antarctic environments. Journal of Basic Microbiology , 51 (6): 590–600, https://doi.org/10.1002/jobm.201000413..

Del Rosso J Q. 2013. Application of protease technology in dermatology: rationale for incorporation into skin care with initial observations on formulations designed for skin cleansing, maintenance of hydration, and restoration of the epidermal permeability barrier. The Journal of Clinical and Aesthetic Dermatology , 6 (6): 14–22..

Dube S, Alam S I, Singh L. 2000. Studies on anaerobic proteolytic bacteria of Leh, India. World Journal of Microbiology and Biotechnology , 16 (3): 297–301, https://doi.org/10.1023/A:1008935425175..

Espósito T S, Amaral I P G, Buarque D S, Oliveira G B, Carvalho Jr L B, Bezerra R S. 2009. Fish processing waste as a source of alkaline proteases for laundry detergent. Food Chemistry , 112 (1): 125–130, https://doi.org/10.1016/j.foodchem.2008.05.049..

Fathi N M, Dileep D, Deepti D. 2005. Potential application of protease isolated from Pseudomonas aeruginosa PD100. Electronic journal of biotechnology , 8 (2): 197–203, https://doi.org/10.2225/vol8-issue2-fulltext-5..

Garciacarreno F L, Dimes L E, Haard N F. 1993. Substrate-gel electrophoresis for composition and molecular weight of proteinases or proteinaceous proteinase inhibitors. Analytical Biochemistry , 214 (1): 65–69, https://doi.org/10.1006/abio.1993.1457..

Gupta R, Beg Q, Lorenz P. 2002. Bacterial alkaline proteases: molecular approaches and industrial applications. Applied Microbiology and Biotechnology , 59 (1): 15–32, https://doi.org/10.1007/s00253-002-0975-y..

Haki G D, Rakshit S K. 2003. Developments in industrially important thermostable enzymes: a review. Bioresource Technology , 89 (1): 17–34, https://doi.org/10.1016/S0960-8524(03)00033-6..

Hassan S W M, Latif H H A E, Ali S M. 2018. Production of cold-active lipase by free and immobilized marine Bacillus cereus HSS: application in wastewater treatment. Frontiers in Microbiology , 9 : 2377, https://doi.org/10.3389/fmicb.2018.02377..

Ichida J M, Krizova L, Lefevre C A, Keener H M, Elwell D L, Burtt Jr E H. 2001. Bacterial inoculum enhances keratin degradation and biofilm formation in poultry compost. Journal of Microbiological Methods , 47 (2): 199–208, https://doi.org/10.1016/S0167-7012(01)00302-5..

Jain D, Pancha I, Mishra S K, Shrivastav A, Mishra S. 2012. Purification and characterization of haloalkaline thermoactive, solvent stable and SDS-induced protease from Bacillus sp. : a potential additive for laundry detergents. Bioresource Technology , 115 : 228–236, https://doi.org/10.1016/j.biortech.2011.10.081..

Joshi S, Satyanarayana T. 2013a. Characteristics and applications of a recombinant alkaline serine protease from a novel bacterium Bacillus lehensis . Bioresource Technology , 131 : 76–85, https://doi.org/10.1016/j.biortech.2012.12.124..

Joshi S, Satyanarayana T. 2013b. Biotechnology of cold-active proteases. Biology , 2 (2): 755–783, https://doi.org/10.3390/biology2020755..

Lee S H, Chung W C, Yu Y J, Rhee Y H. 2009. Effect of alkaline protease-producing Exiguobacterium sp. YS1 inoculation on the solubilization and bacterial community of waste activated sludge. Bioresource Technology , 100 (20): 4597–4603, https://doi.org/10.1016/j.biortech.2009.04.056..

Li P Y, Zhang Y Q, Zhang Y, Jiang W X, Wang Y J, Zhang Y S, Sun Z Z, Li C Y, Zhang Y Z, Shi M, Song X Y, Zhao L S, Chen X L. 2020a. Study on a novel cold-active and halotolerant monoacylglycerol lipase widespread in marine bacteria reveals a new group of bacterial monoacylglycerol lipases containing unusual C(A/S) HSMG catalytic motifs. Frontiers in Microbiology , 11 : 9, https://doi.org/10.3389/fmicb.2020.00009..

Li Q, Yi L, Marek P, Iverson B L. 2013. Commercial proteases: present and future. FEBS Letters , 587 (8): 1155–1163, https://doi.org/10.1016/j.febslet.2012.12.019.

Li X P, Zhang Y, Wu T, Sun X M, Yang T, Wang L J, Li X Y, Wang J Z, Wang Y H, Yu H T. 2020b. Major ions in drinking and surface waters from five cities in arid and semi-arid areas, NW China: spatial occurrence, water chemistry, and potential anthropogenic inputs. Environmental Science and Pollution Research , 27 (5): 5456–5468, https://doi.org/10.1007/s11356-019-07149-9..

Ma C L, Ni X M, Chi Z M, Ma L Y, Gao L M. 2007. Purification and characterization of an alkaline protease from the marine yeast Aureobasidium pullulans for bioactive peptide production from different sources. Marine Biotechnology , 9 (3): 343–351, https://doi.org/10.1007/s10126-006-6105-6..

Makino K, Koshikawa T, Nishihara T, Ichikawa T, Kondo M. 1981. Studies on protease from marine bacteria. 1 Isolation of marine Pseudomonas sp. 145–2 and purification of protease. Microbios , 31 (124): 103–112..

Mamiatis T, Fritsch E F, Sambrook J, Engel J. 1985. Molecular cloning—A laboratory manual. New York: Cold Spring Harbor Laboratory. 1982, 545 S., 42 ＄. Acta Biotechnologica , 5 (1): 104, https://doi.org/10.1002/abio.370050118..

Martin M, Biver S, Steels S, Barbeyron T, Jam M, Portetelle D, Michel G, Vandenbol M. 2014. Identification and characterization of a halotolerant, cold-active marine endo-β-1, 4-glucanase by using functional metagenomics of seaweed-associated microbiota. Applied and Environmental Microbiology , 80 (16): 4958–4967, https://doi.org/10.1128/AEM.01194-14..

Marx J C, Collins T, D'Amico S, Feller G, Gerday C. 2007. Cold-adapted enzymes from marine antarctic microorganisms. Marine Biotechnology , 9 (3): 293–304, https://doi.org/10.1007/s10126-006-6103-8..

Niyonzima F N, More S S. 2015a. Coproduction of detergent compatible bacterial enzymes and stain removal evaluation. Journal of Basic Microbiology , 55 (10): 1149–1158, https://doi.org/10.1002/jobm.201500112..

Niyonzima F N, More S. 2015b. Detergent-compatible proteases: microbial production, properties, and stain removal analysis. Preparative Biochemistry & Biotechnology , 45 (3): 233–258, https://doi.org/10.1080/10826068.2014.907183..

Oberoi R, Beg Q K, Puri S, Saxena R K, Gupta R. 2001. Characterization and wash performance analysis of an SDS-stable alkaline protease from a Bacillus sp. World Journal of Microbiology and Biotechnology , 17 (5): 493–497, https://doi.org/10.1023/A:1011911109179..

Ogino H, Ishikawa H. 2001. Enzymes which are stable in the presence of organic solvents. Journal of Bioscience and Bioengineering , 91 (2): 109–116, https://doi.org/10.1016/S1389-1723(01)80051-7..

Oh K H, Seong C S, Lee S W, Kwon O S, Park Y S. 1999. Isolation of a psychrotrophic Azospirillum sp. and characterization of its extracellular protease. FEMS Microbiology Letters , 174 (1): 173–178, https://doi.org/10.1111/j.1574-6968.1999.tb13565.x..

Outtrup H, Boyce C O L. 1990. Microbial proteinases and biotechnology. In : Fogarty W M, Kelly C T eds. Microbial Enzymes and Biotechnology. Springer, Dordrecht. p. 227–254, https://doi.org/10.1007/978-94-009-0765-2_6 https://doi.org/10.1007/978-94-009-0765-2_6 ..

Sun M, Wang Y J, Zhang Y B, Hong Y G, Yan X L, Chen Y Z, Wang C B, Liu X P. 2000. Study on isolation and culture condition of one marine bacterium YS-9412-130 producing low-temperature alkaline protease. Marine Fisheries Research , 21 (4): 1–5. (in Chinese with English abstract).

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution , 30 (12): 2725–2729, https://doi.org/10.1093/molbev/mst197..

Ting L, Williams T J, Cowley M J, Lauro F M, Guilhaus M, Raftery M J, Cavicchioli R. 2010. Cold adaptation in the marine bacterium, Sphingopyxis alaskensis , assessed using quantitative proteomics. Environmental Microbiology , 12 (10): 2658–2676, https://doi.org/10.1111/j.1462-2920.2010.02235.x..

Vázquez S C, Hernández E, Cormack W P M. 2008.: Extracellular proteases from the Antarctic marine Pseudoalteromonas sp. P96-47 strain. Revista Argentina de Microbiologia , 40 (1): 63–71, https://doi.org/10.1016/10.resmic.2007.12.001..

Wang F, Hao J H, Yang C Y, Sun M. 2010. Cloning, expression, and identification of a novel extracellular cold-adapted alkaline protease gene of the marine bacterium strain YS-80–122. Applied Biochemistry and Biotechnology , 162 (5): 1497–1505, https://doi.org/10.1007/s12010-010-8927-y..

Wu G B, Zhang X N, Wei L, Wu G J, Kumar A, Mao T, Liu Z D. 2015a. A cold-adapted, solvent and salt tolerant esterase from marine bacterium Psychrobacter pacificensis . International Journal of Bologcal Macromolecules , 81 : 180–187, https://doi.org/10.1016/j.ijbiomac.2015.07.045..

Wu S M, Liu G, Zhang D C, Li C X, Sun C M. 2015b. Purification and biochemical characterization of an alkaline protease from marine bacteria Pseudoalteromonas sp. 129–1. Journal of Basic Microbiology , 55 (12): 1427–1434, https://doi.org/10.1002/jobm.201500327..

Xu J X, Zhuang Y, Wu B, Su L, He B F. 2013. Calcium-ion-induced stabilization of the protease from Bacillus cereus WQ9-2 in aqueous hydrophilic solvents: effect of calcium ion binding on the hydration shell and intramolecular interactions. Journal of Biological Inorganic Chemistry , 18 (2): 211–221, https://doi.org/10.1007/s00775-012-0966-0..

Zhou M Y, Chen X L, Zhao H L, Dang H Y, Luan X W, Zhang X Y, He H L, Zhou B C, Zhang Y Z. 2009. Diversity of both the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea. Microbial Ecology , 58 (3): 582–590, https://doi.org/10.1007/s00248-009-9506-z..

Zhou Y, Chen X H, Li X, Han Y T, Wang Y N, Yao R Y, Li S Y 2019. Purification and characterization of a new cold-adapted and thermo-tolerant chitosanase from marine bacterium Pseudoalteromonas sp. SY39. Molecules , 24 (1): 183, https://doi.org/10.3390/molecules24010183..

浏览量

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

工具集

关联资源

Purification and characterization of lectin from humoral fluids ofCharybdis feriatus

Purification and characterization of vitellin in mature ovary of marine crab Charybdis japonica

Purification and characterization of novel κ-carrageenase from marine Tamlana sp. HC4

Characterization of proteases from Planomicrobium sp. L-2 isolated from the gastrointestinal tract of Octopus variabilis (Sasaki)

Characterization and stability of sedimentary colloids in different ecology regions in Taihu Lake