Partial Characterization of Hydrolytic Enzymes Produced by Bacillus Strains Isolated from Balıklıgöl, Turkey
Öz
In the present study, forty-five bacterial isolates were obtained from previously unstudied soil samples in Balıklıgöl, Şanlıurfa. Based on their enzim production capacities, six bacterial isolates designated as BGL-22, BGL-26, BGL-27, BGL-37, BGL-38 and BGL-39 were selected for further studies. Conventional and molecular identification results showed that the bacteria belonged to Bacillus genus. Among these strains, the highest activities for amylase (11.44 U mL-1), lipase (1.12 U mL-1) and protease (2.61 U mL-1) were determined for Bacillus sp. BGL-37. Enzymatic characterization studies demonstrated that the activities of acid-stable amylase and alkaline-stable lipase remained unchanged up to 50℃, while alkaline-protease was retained about 90% of its activity up to 40℃. The findings suggested that these enzymes providing environmentally compatible processes under relatively mild conditions have potential to be used in several fields such as food processing and detergent industry.
Anahtar Kelimeler
Bacillus sp. Extracellular enzymes Characterization 16S rRNA sequencing
Destekleyen Kurum
the Scientific Research Unit of Harran University
Proje Numarası
Project No: HUBAK-12007
Teşekkür
This study was supported by the Scientific Research Unit of Harran University (Project No: HUBAK-12007). We also thank MedSanTek for performing sequence analysis of PCR products.
Kaynakça
- Adrio JL, Demain AL 2014. Microbial enzymes: tools for biotechnological processes. Biomolecules 4: 117-139.
- Bergey DH, Holt JG 1994. Bergey’s manual of determinative microbiology, 9th edn. Williams and Wilkins, Baltimore.
- Bhunia B, Dutta D, Chaudhur S 2011. Extracellular alkaline protease from Bacillus licheniformis NCIM-2042: improving enzyme activity assay and characterization. Eng Life Sci 11: 207-215.
- Bradford MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
- Chapman J, Ismail AE, Dinu CZ 2018. Industrial applications of enzymes: recent advances, techniques, and outlooks. Catalysts 8: 238.
- Dahiya P, Rathi B 2015. Characterization and application of alkaline α-amylase from Bacillus licheniformis MTCC1483 as a detergent additive. Int Food Res J 22: 1293-1297.
- Deljou A, Arezi I 2016. Production of thermostable extracellular a-amylase by a moderate thermophilic Bacillus licheniformis-AZ2 isolated from Qinarje Hot spring (Ardebil prov. of Iran). Period Biol 118: 405-416.
- Divakaran D, Chandran A, Chandran P 2011. Comparative study on production of α-amylase from Bacillus licheniformis strains. Braz J Microbiol 42: 1397-1404.
- El Hadj-Ali N, Agrebi R, Ghorbel-Frikha B, Sellami-Kamoun A, Kanoun S, Nasri M 2007. Biochemical and molecular characterization of a detergent stable alkaline serine-protease from a newly isolated Bacillus licheniformis NH1. Enzyme Microb Technol 40: 515-523.
- Felsenstein J 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39: 783-791.
- Gomaa EZ 2013. Optimization and characterization of alkaline protease and carboxymethyl-cellulase produced by Bacillus pumillus grown on Ficus nitida wastes. Braz J Microbiol 44: 529-537.
- Jamrath T, Lindner C, Popovic MK, Bajpai R 2012. Production of amylases and proteases by Bacillus caldolyticus from food industry wastes. Food Technol Biotech 50: 355-361.
- Kumar S, Kikon K, Upadhyay A, Kanwar SS, Gupta R 2005. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. Protein Expr Purif 41: 38-44.
- Kumar S, Stecher G, Tamura K 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33: 1870-1874.
- Mahdavi A, Sajedi, RH, Rassa M, Jafarian V 2010. Characterization of an α-amylase with broad temperature activity from an acid-neutralizing Bacillus cereus strain. Iran J Biotechnol 8: 103-111.
- Mehrotra S, Pandey PK, Gaur R, Darmwa NS 1999. The production of alkaline protease by a Bacillus species isolate. Bioresour Technol 67: 201-203.
- Nei M, Kumar S 2000. Molecular evolution and phylogenetics. Oxford University Press, New York. Parrado J, Rodriguez-Morgado B, Tejada M, Hernandez T, Garcia C 2014. Proteomic analysis of enzyme production by Bacillus licheniformis using different feather wastes as the sole fermentation media. Enzyme Microb Technol 57: 1-7.
- Prasad S, Roy I 2018. Converting enzymes into tools of industrial importance. Recent Pat Biotechnol 12: 33-56.
- Rapp P, Backhaus S 1992. Formation of extracellular lipases by filamentous fungi, yeasts, and bacteria. Enzyme Microb Technol 14: 938-943.
- Rastall R 2007. Novel enzyme technology for food applications. Woodhead Publishing, Cambridge.
- Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, Rebello S, Pandey A 2018. Applications of microbial enzymes in food industry. Food Technol Biotech 56: 16-30.
- Ray RC, Rosell CM 2017. Microbial enzyme technology in food applications. CRC Press, Boca Raton. Rick W, Stegbauer HP 1974. α-amylase measurement of reducing group (Methods of enzymatic analysis, Academic Press, New York: Ed. Bergmeyer HU) 885-890.
- Saitou N, Nei M 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406-425.
- Sanchez S, Demain A 2017. Useful microbial enzymes-an introduction (Biotechnology of microbial enzymes: production, biocatalysis and industrial applications, Elsevier Academic Press, Amsterdam: Ed. Brahmachari G, Demain AL, Adrio JL) 1-11.
- Saraswat R, Verma V, Sistla S, Bhushan I 2017. Evaluation of alkali and thermotolerant lipase from an indigenous isolated Bacillus strain for detergent formulation. Electron J Biotechn 30: 33-38.
- Schallmey M, Singh A, Ward OP 2004. Developments in the use of Bacillus species for industrial production. Can J Microbiol 50: 1-7.
- Sharma D, Kumbhar BK, Verma AK, Tewari L 2014. Optimization of critical growth parameters for enhancing extracellular lipase production by alkalophilic Bacillus sp. Biocatal Agric Biotechnol 3: 205-211.
- Sharma R, Soni SK, Vohra RM, Gupta LK, Gupta JK 2002. Purification and characterization of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1. Process Biochem 37: 1075-1084.
- Yilmaz B, Baltaci MO, Sisecioglu M, Adiguzel A 2016. Thermotolerant alkaline protease enzyme from Bacillus licheniformis A10: purification, characterization, effects of surfactants and organic solvents. J Enzyme Inhib Med Chem 31: 1241-1247.
Balıklıgöl, Türkiye’den İzole Edilen Bacillus Suşları Tarafından Üretilen Hidrolitik Enzimlerin Kısmi Karakterizasyonu
Öz
Bu çalışmada, Balıklıgöl, Şanlıurfa’dan elde edilmiş ve daha önceden çalışılmamış toprak örneklerinden kırk beş bakteriyel izolat elde edilmiştir. Enzim üretim kapasitelerine göre bu izolatlar arasından BGL-22, BGL-26, BGL-27, BGL-37, BGL-38 ve BGL-39 olarak isimlendirilen altı bakteriyal izolat ileriki denemeler için seçilmiştir. Konvansiyonel ve moleküler tanılama sonuçları, bu bakterilerin Bacillus cinsine dahil olduğunu göstermiştir. Bu suşlar arasından Bacillus sp. BGL-37, amilaz (11.44 U mL-1), lipaz (1.12 U mL-1) ve proteaz (2.61 U mL-1) açısından en yüksek aktiviteyi göstermiştir. Enzimatik karakterizasyon çalışmaları, alkali proteazın 40ºC’ye kadar yaklaşık %90 aktivitesini koruduğunu, asit-kararlı amilaz ve alkali-kararlı lipazın ise 50ºC’ye kadar aktivitelerinde herhangi bir düşüş yaşanmadan aktivite gösterdiklerini ortaya koymuştur. Elde edilen bulgular, nispeten ılımlı koşullar altında çevresel olarak uyumlu süreçlerin gerçekleşmesini sağlayan bu enzimlerin, gıdaların işlenmesi ve deterjan endüstrisi gibi çeşitli alanlarda kullanılma potansiyeline sahip olduğunu göstermiştir.
Anahtar Kelimeler
Bacillus sp. Hücre dışı enzimler Karakterizasyon 16S rRNA dizileme
Proje Numarası
Project No: HUBAK-12007
Kaynakça
- Adrio JL, Demain AL 2014. Microbial enzymes: tools for biotechnological processes. Biomolecules 4: 117-139.
- Bergey DH, Holt JG 1994. Bergey’s manual of determinative microbiology, 9th edn. Williams and Wilkins, Baltimore.
- Bhunia B, Dutta D, Chaudhur S 2011. Extracellular alkaline protease from Bacillus licheniformis NCIM-2042: improving enzyme activity assay and characterization. Eng Life Sci 11: 207-215.
- Bradford MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
- Chapman J, Ismail AE, Dinu CZ 2018. Industrial applications of enzymes: recent advances, techniques, and outlooks. Catalysts 8: 238.
- Dahiya P, Rathi B 2015. Characterization and application of alkaline α-amylase from Bacillus licheniformis MTCC1483 as a detergent additive. Int Food Res J 22: 1293-1297.
- Deljou A, Arezi I 2016. Production of thermostable extracellular a-amylase by a moderate thermophilic Bacillus licheniformis-AZ2 isolated from Qinarje Hot spring (Ardebil prov. of Iran). Period Biol 118: 405-416.
- Divakaran D, Chandran A, Chandran P 2011. Comparative study on production of α-amylase from Bacillus licheniformis strains. Braz J Microbiol 42: 1397-1404.
- El Hadj-Ali N, Agrebi R, Ghorbel-Frikha B, Sellami-Kamoun A, Kanoun S, Nasri M 2007. Biochemical and molecular characterization of a detergent stable alkaline serine-protease from a newly isolated Bacillus licheniformis NH1. Enzyme Microb Technol 40: 515-523.
- Felsenstein J 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39: 783-791.
- Gomaa EZ 2013. Optimization and characterization of alkaline protease and carboxymethyl-cellulase produced by Bacillus pumillus grown on Ficus nitida wastes. Braz J Microbiol 44: 529-537.
- Jamrath T, Lindner C, Popovic MK, Bajpai R 2012. Production of amylases and proteases by Bacillus caldolyticus from food industry wastes. Food Technol Biotech 50: 355-361.
- Kumar S, Kikon K, Upadhyay A, Kanwar SS, Gupta R 2005. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. Protein Expr Purif 41: 38-44.
- Kumar S, Stecher G, Tamura K 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33: 1870-1874.
- Mahdavi A, Sajedi, RH, Rassa M, Jafarian V 2010. Characterization of an α-amylase with broad temperature activity from an acid-neutralizing Bacillus cereus strain. Iran J Biotechnol 8: 103-111.
- Mehrotra S, Pandey PK, Gaur R, Darmwa NS 1999. The production of alkaline protease by a Bacillus species isolate. Bioresour Technol 67: 201-203.
- Nei M, Kumar S 2000. Molecular evolution and phylogenetics. Oxford University Press, New York. Parrado J, Rodriguez-Morgado B, Tejada M, Hernandez T, Garcia C 2014. Proteomic analysis of enzyme production by Bacillus licheniformis using different feather wastes as the sole fermentation media. Enzyme Microb Technol 57: 1-7.
- Prasad S, Roy I 2018. Converting enzymes into tools of industrial importance. Recent Pat Biotechnol 12: 33-56.
- Rapp P, Backhaus S 1992. Formation of extracellular lipases by filamentous fungi, yeasts, and bacteria. Enzyme Microb Technol 14: 938-943.
- Rastall R 2007. Novel enzyme technology for food applications. Woodhead Publishing, Cambridge.
- Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, Rebello S, Pandey A 2018. Applications of microbial enzymes in food industry. Food Technol Biotech 56: 16-30.
- Ray RC, Rosell CM 2017. Microbial enzyme technology in food applications. CRC Press, Boca Raton. Rick W, Stegbauer HP 1974. α-amylase measurement of reducing group (Methods of enzymatic analysis, Academic Press, New York: Ed. Bergmeyer HU) 885-890.
- Saitou N, Nei M 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406-425.
- Sanchez S, Demain A 2017. Useful microbial enzymes-an introduction (Biotechnology of microbial enzymes: production, biocatalysis and industrial applications, Elsevier Academic Press, Amsterdam: Ed. Brahmachari G, Demain AL, Adrio JL) 1-11.
- Saraswat R, Verma V, Sistla S, Bhushan I 2017. Evaluation of alkali and thermotolerant lipase from an indigenous isolated Bacillus strain for detergent formulation. Electron J Biotechn 30: 33-38.
- Schallmey M, Singh A, Ward OP 2004. Developments in the use of Bacillus species for industrial production. Can J Microbiol 50: 1-7.
- Sharma D, Kumbhar BK, Verma AK, Tewari L 2014. Optimization of critical growth parameters for enhancing extracellular lipase production by alkalophilic Bacillus sp. Biocatal Agric Biotechnol 3: 205-211.
- Sharma R, Soni SK, Vohra RM, Gupta LK, Gupta JK 2002. Purification and characterization of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1. Process Biochem 37: 1075-1084.
- Yilmaz B, Baltaci MO, Sisecioglu M, Adiguzel A 2016. Thermotolerant alkaline protease enzyme from Bacillus licheniformis A10: purification, characterization, effects of surfactants and organic solvents. J Enzyme Inhib Med Chem 31: 1241-1247.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Yapısal Biyoloji |
| Bölüm | ARAŞTIRMA MAKALESİ (Research Article) |
| Yazarlar | |
| Proje Numarası | Project No: HUBAK-12007 |
| Yayımlanma Tarihi | 31 Ağustos 2021 |
| Gönderilme Tarihi | 12 Ekim 2020 |
| Kabul Tarihi | 3 Aralık 2020 |
| Yayımlandığı Sayı | Yıl 2021Cilt: 24 Sayı: 4 |
2022-JIF = 0.500
2022-JCI = 0.170
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