Saccharification of Hazelnut and Rhododendron Biomasses Using β-xylanase from Thermotoga naphthophila
Abstract
Enzymes can be used in various biotechnological applications due to the easy and cheap production. Since xylanase enzymes are preferred in various industries, researchon this enzyme is extensively being carried out. In this study, the β-xylanase gene was cloned from Thermotoga naphthophila, a thermophilic organism. The expression vector pET21a(+) was expressed in Escherichia coli BL21 (DE3). As a result of the studies, the pH, temperature and IPTG concentration of the enzyme were optimized to obtain highest expression. Dinitrosalicylic acid (DNS) was used to determine sugar content of the enzyme. The molecular mass of the
purified β-xylanase enzyme was determined using sodium dodecyl sulfate (SDS) polyacrylamide gel
electrophoresis. The molecular mass of the enzyme was calculated to be 38 kDa. Enzymatic hydrolysis of
hazelnut shell, rhododendron branch and rhododendron leaves was performed. Released reducing sugar contents from the enzymatic hydrolysis were calculated as 0.8461 mg mL-1, 0.6976 mg mL-1 and 0.3605 mg mL-1 for hazelnut shell, rhododendron branch, and rhododendron leaf respectively. In conclusion, β-xylanase enzyme can be an effective source for enzymatic hydrolysis to produce fermentable sugars from such biomasses.
Keywords
Thanks
This project was supported by PAK-Turk Collaborative Research Program of Turkish Council of Higher Education (YÖK).
References
- Arslan Y, Saraçoğlu NE, 2010. Effects of pretreatment methods for hazelnut shell hydrolysate fermentation with Pichia Stipitis to ethanol. Bioresource Technology, 101: 8664-8670.
- Dodd D, Cann, IKO, 2009. Enzymatic deconstruction of xylan for biofuel production. GCB Bioenergy, 1(1): 2-17.
- Frock AD, Notey JS, Kelly RM, 2010. The genus Thermotoga: recent developments. Environmental Technology, 31(10): 1169-1181.
- Hamid A, Aftab MN, 2019. Cloning, purification, and caharacterization of recombinant thermostable β-xylanase Tnap_0700 from Thermotoga naphthophila. Applied Biochemistry and Biotechnology, 189(4): 1274-1290.
- Haq I, Hussain Z, Khan MA, Muneer B, Afzal S, Majeed S, Akram F, 2012. Kinetic and thermodynamic study of cloned thermostable endo-1, 4-β-xylanase from Thermotoga petrophila in mesophilic host. Molecular biology reports, 39(7): 7251-7261.
- Hoşgün EZ, Berikten D, Kıvanç M, Bozan B, 2017. Ethanol production from hazelnut shells through enzymatic saccharification and fermantion by low-temperature alkali pretreatment. Fuel, 196: 280-278.
- Jegannathan KR, Nielsen PH, 2013. Environmental assessment of enzyme use in industrial production – a literature review. Journal of Cleaner Production, 42: 228-240.
- López, L, Rivas S, Moure A, Vila C, Parajó JC, 2020. Development of pretreatment strategies for the fractionation of hazelnut shells in the scope of biorefinery. Agronomy, 10(10): 1568.
- Luo J, Fang Z, Smith Jr RL, 2014. Ultrasound-enhanced conversion of biomass to biofuels. Progress in Energy and Combustion Science, 41: 56-93.
- Miller GL, 1959. Use of dinitrosalicylic asid reagent for determination of reducing sugar. Analytical Chemistry, 31(3): 426-428.
- Özbucak TB, Türkiş S, Çakmak A, 2009. Ordu çevresinde yayılış gösteren bazı rhododendron türleri üzerine ekolojil bir çalışma. Biyoloji Bilimleri Araştırma Dergisi, 2(2): 71-77.
- Phitsuwan P, Sakka K, Ratanakhanokchai K, 2016. Structural changes and enzymatic response of Napier grass (Pennisetum purpureum) stem induced by alkaline pretreatment. Bioresourse Technology, 218: 247-256.
- Pinar O, Karaosmanoğlu K, Sayar NA, Kula C, Kazan D, Sayar AA, 2017. Assessment of hazelnut husk as a lignocellulosic feedstock for the production of fermentable sugars and lignocellulolytic enzymes. 3 Biotech, 7(6): 367.
- Rani S, Nand K, 1996. Development of cellulase-free xylanase-producing anaerobic consortia for use the use of lignocellulosic wastes. Enzyme and Microbial Technology 18(1): 23-28.
- Shi H, Zhang Y, Li X, Huang Y, Wang L, Wang, Y, Wang F, 2013. A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization. Biotechnology for biofuels, 6(1): 26.
- Shi H, Zhang, Y, Zhong H, Huang Y, Li X, Wang F, 2014. Cloning over-expression and characterization of a thermo-tolerant xylanase from Thermotoga thermarum. Biotechnology Letters, 36(3): 587-593.
- Simpson HD, Haufler UR, Daniel RM, 1991. An extremely thermostable xylanase from the thermophilic eubacterium Thermotoga. Biochemical Journal, 277(2): 413-417.
- Singh J, Suhag M, Dhaka A, 2015. Augmented digestion of lignocellulose by steam explosion, acid and alkaline pretreatment methods: areview. Carbohydrate polymers, 117: 624-631.
- Sun Y, Cheng J, 2002. Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresource Technology, 83(1): 1-11.
- Topal Ş, 1985. Enzimler, Mikrobiyolojik Yolla Enzim Üretimi ve Bu Teknolojide Renni’nin Yeri. Tubitak Marmara Araştırma Enstitüsü, Gebze/Kocaeli, 26 s.
- Zafar A, Aftab MN, ud Din Z, Aftab S, Iqbal I, Shahid A, ul Haq I, 2016. Cloning, expression and purification of xylanase gene from Bacillus licheniformis for use in saccharification of plant biomass. Applied Biochemistry and Biotechnology, 178(2): 294-311.
- Zhou J, Shi P, Zhang R, Huang H, Meng K, Yang P, Yao B, 2011. Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease-and SDS-resistant xylanase. Journal of Industrial Microbiology and Biotechnology, 38(4): 523-530
- Zverlov V, Piotukh K, Dakhova O, Velikodvorskaya G, Borriss R, 1996. The multidomain xylanase A of the hyperthermophilic bacterium Thermotoga neapolitana is extremely thermoresistant. Applied Microbiology and Biotechnology, 45(1-2): 245-247.
Saccharification of Hazelnut and Rhododendron Biomasses Using β-xylanase from Thermotoga naphthophila
Abstract
Enzymes can be used in various biotechnological applications due to the easy and cheap production. Since xylanase enzymes are preferred in various industries, researchon this enzyme is extensively being carried out. In this study, the β-xylanase gene was cloned from Thermotoga naphthophila, a thermophilic organism. The expression vector pET21a(+) was expressed in Escherichia coli BL21 (DE3). As a result of the studies, the pH, temperature and IPTG concentration of the enzyme were optimized to obtain highest expression. Dinitrosalicylic acid (DNS) was used to determine sugar content of the enzyme. The molecular mass of the
purified β-xylanase enzyme was determined using sodium dodecyl sulfate (SDS) polyacrylamide gel
electrophoresis. The molecular mass of the enzyme was calculated to be 38 kDa. Enzymatic hydrolysis of
hazelnut shell, rhododendron branch and rhododendron leaves was performed. Released reducing sugar contents from the enzymatic hydrolysis were calculated as 0.8461 mg mL-1, 0.6976 mg mL-1 and 0.3605 mg mL-1 for hazelnut shell, rhododendron branch, and rhododendron leaf respectively. In conclusion, β-xylanase enzyme can be an effective source for enzymatic hydrolysis to produce fermentable sugars from such biomasses.
References
- Arslan Y, Saraçoğlu NE, 2010. Effects of pretreatment methods for hazelnut shell hydrolysate fermentation with Pichia Stipitis to ethanol. Bioresource Technology, 101: 8664-8670.
- Dodd D, Cann, IKO, 2009. Enzymatic deconstruction of xylan for biofuel production. GCB Bioenergy, 1(1): 2-17.
- Frock AD, Notey JS, Kelly RM, 2010. The genus Thermotoga: recent developments. Environmental Technology, 31(10): 1169-1181.
- Hamid A, Aftab MN, 2019. Cloning, purification, and caharacterization of recombinant thermostable β-xylanase Tnap_0700 from Thermotoga naphthophila. Applied Biochemistry and Biotechnology, 189(4): 1274-1290.
- Haq I, Hussain Z, Khan MA, Muneer B, Afzal S, Majeed S, Akram F, 2012. Kinetic and thermodynamic study of cloned thermostable endo-1, 4-β-xylanase from Thermotoga petrophila in mesophilic host. Molecular biology reports, 39(7): 7251-7261.
- Hoşgün EZ, Berikten D, Kıvanç M, Bozan B, 2017. Ethanol production from hazelnut shells through enzymatic saccharification and fermantion by low-temperature alkali pretreatment. Fuel, 196: 280-278.
- Jegannathan KR, Nielsen PH, 2013. Environmental assessment of enzyme use in industrial production – a literature review. Journal of Cleaner Production, 42: 228-240.
- López, L, Rivas S, Moure A, Vila C, Parajó JC, 2020. Development of pretreatment strategies for the fractionation of hazelnut shells in the scope of biorefinery. Agronomy, 10(10): 1568.
- Luo J, Fang Z, Smith Jr RL, 2014. Ultrasound-enhanced conversion of biomass to biofuels. Progress in Energy and Combustion Science, 41: 56-93.
- Miller GL, 1959. Use of dinitrosalicylic asid reagent for determination of reducing sugar. Analytical Chemistry, 31(3): 426-428.
- Özbucak TB, Türkiş S, Çakmak A, 2009. Ordu çevresinde yayılış gösteren bazı rhododendron türleri üzerine ekolojil bir çalışma. Biyoloji Bilimleri Araştırma Dergisi, 2(2): 71-77.
- Phitsuwan P, Sakka K, Ratanakhanokchai K, 2016. Structural changes and enzymatic response of Napier grass (Pennisetum purpureum) stem induced by alkaline pretreatment. Bioresourse Technology, 218: 247-256.
- Pinar O, Karaosmanoğlu K, Sayar NA, Kula C, Kazan D, Sayar AA, 2017. Assessment of hazelnut husk as a lignocellulosic feedstock for the production of fermentable sugars and lignocellulolytic enzymes. 3 Biotech, 7(6): 367.
- Rani S, Nand K, 1996. Development of cellulase-free xylanase-producing anaerobic consortia for use the use of lignocellulosic wastes. Enzyme and Microbial Technology 18(1): 23-28.
- Shi H, Zhang Y, Li X, Huang Y, Wang L, Wang, Y, Wang F, 2013. A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization. Biotechnology for biofuels, 6(1): 26.
- Shi H, Zhang, Y, Zhong H, Huang Y, Li X, Wang F, 2014. Cloning over-expression and characterization of a thermo-tolerant xylanase from Thermotoga thermarum. Biotechnology Letters, 36(3): 587-593.
- Simpson HD, Haufler UR, Daniel RM, 1991. An extremely thermostable xylanase from the thermophilic eubacterium Thermotoga. Biochemical Journal, 277(2): 413-417.
- Singh J, Suhag M, Dhaka A, 2015. Augmented digestion of lignocellulose by steam explosion, acid and alkaline pretreatment methods: areview. Carbohydrate polymers, 117: 624-631.
- Sun Y, Cheng J, 2002. Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresource Technology, 83(1): 1-11.
- Topal Ş, 1985. Enzimler, Mikrobiyolojik Yolla Enzim Üretimi ve Bu Teknolojide Renni’nin Yeri. Tubitak Marmara Araştırma Enstitüsü, Gebze/Kocaeli, 26 s.
- Zafar A, Aftab MN, ud Din Z, Aftab S, Iqbal I, Shahid A, ul Haq I, 2016. Cloning, expression and purification of xylanase gene from Bacillus licheniformis for use in saccharification of plant biomass. Applied Biochemistry and Biotechnology, 178(2): 294-311.
- Zhou J, Shi P, Zhang R, Huang H, Meng K, Yang P, Yao B, 2011. Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease-and SDS-resistant xylanase. Journal of Industrial Microbiology and Biotechnology, 38(4): 523-530
- Zverlov V, Piotukh K, Dakhova O, Velikodvorskaya G, Borriss R, 1996. The multidomain xylanase A of the hyperthermophilic bacterium Thermotoga neapolitana is extremely thermoresistant. Applied Microbiology and Biotechnology, 45(1-2): 245-247.
Details
| Primary Language | English |
|---|---|
| Subjects | Chemical Engineering |
| Journal Section | Kimya / Chemistry |
| Authors | |
| Publication Date | June 1, 2021 |
| Submission Date | November 17, 2020 |
| Acceptance Date | January 25, 2021 |
| Published in Issue | Year 2021 Volume: 11 Issue: 2 |