Rational Development Strategies with N-Glycosylation Engineering of Rhizomucor miehei Lipase for Biodiesel Production

2021 ◽  
Author(s):  
Miao Tian ◽  
Zhiyuan Wang ◽  
Junying Fu ◽  
Pengmei Lv ◽  
Cuiyi Liang ◽  
...  
Keyword(s):  
Rhizomucor Miehei ◽  
Development Strategies ◽  
Biodiesel Production ◽  
Rational Development ◽  
Glycosylation Engineering

Rapid and high-density covalent immobilization of Rhizomucor miehei lipase using a multi component reaction: application in biodiesel production

RSC Advances ◽  
2015 ◽  
Vol 5 (41) ◽  
pp. 32698-32705 ◽  
Author(s):  
Mehdi Mohammadi ◽  
Maryam Ashjari ◽  
Shaghayegh Dezvarei ◽  
Maryam Yousefi ◽  
Mohadese Babaki ◽  
...  
Keyword(s):  
Mild Condition ◽  
High Capacity ◽  
Rhizomucor Miehei ◽  
Covalent Immobilization ◽  
Ugi Reaction ◽  
High Density ◽  
Biodiesel Production ◽  
Rhizomucor Miehei Lipase ◽  
Miehei Lipase

Rapid and high capacity immobilization of Rhizomucor miehei lipase on aldehyde-functionalized supports was performed under mild condition via a multi component reaction. The mechanism of immobilization reaction was determined as the Ugi reaction.

Application of Rhizomucor miehei lipase-displaying Pichia pastoris whole cell for biodiesel production using agro-industrial residuals as substrate

2021 ◽  
Vol 189 ◽  
pp. 734-743
Author(s):  
Raphael Oliveira Sena ◽  
Candida Carneiro ◽  
Marcelo Victor Holanda Moura ◽  
Gabriela Coelho Brêda ◽  
Martina C.C. Pinto ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Rhizomucor Miehei ◽  
Biodiesel Production ◽  
Rhizomucor Miehei Lipase ◽  
Whole Cell ◽  
Miehei Lipase

scholarly journals Magnetic COFs as satisfied support for lipase immobilization and recovery to effectively achieve the production of biodiesel by great maintenance of enzyme activity.

2020 ◽  
Author(s):  
Zi-Wen Zhou ◽  
Xiu Xing ◽  
Jun Li ◽  
Zu-E Hu ◽  
Zong-Bo Xie ◽  
...  
Keyword(s):  
Surface Area ◽  
High Surface ◽  
Enzymatic Catalysis ◽  
High Surface Area ◽  
Rhizomucor Miehei ◽  
Catalytic Performance ◽  
Biodiesel Production ◽  
Core Shell ◽  
Water Medium ◽  
Practical Applications

Abstract Background: Production of biodiesel from renewable sources such as non-edible vegetable oils by enzymatic catalysis has been a hotspot but remains a challenge on the efficient use of an enzyme. COFs featuring large surface area and porosity should be applied as an ideal support to impede aggregation of lipase and methanol. However, the naturally low density limits its application. In this work, we reported a facile synthesis of core-shell magnetic COF composite (Fe3O4@COF-OMe) to immobilize RML (Rhizomucor miehei lipase), in order to achieve its utilization in biodiesel production.Result: This strategy gives extrinsic magnetic property, and the magnetic COFs is much heavier and could disperse in water medium well, facilitating the attachment with enzyme. The resultant biocomposite exhibited an excellent capacity of RML due to its high surface area and fast response to the external magnetic field, as well as good chemical stability. The core-shell magnetic COF-OMe structure not only achieved highly efficient immobilization and recovery processes, but also maintained the activity of lipase to a great extent. RML@Fe3O4@COF-OMe performed well in practical applications, while free lipase did not. The biocomposite successfully achieved the production of biodiesel from Jatropha curcas Oil with a yield of about 70% in the optimized conditions. Conclusion: Magnetic COFs (Fe3O4@COF-OMe) for RML immobilization greatly improved catalytic performance in template reaction and biodiesel preparation. The magneticity makes it easily recovered and separated from system. This first successful attempt of COFs-based immobilized enzyme broadened the prospect of biodiesel production by COFs with some inspiration.

scholarly journals Magnetic COFs as satisfied support for lipase immobilization and recovery to effectively achieve the production of biodiesel by great maintenance of enzyme activity

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Zi-Wen Zhou ◽  
Chun-Xian Cai ◽  
Xiu Xing ◽  
Jun Li ◽  
Zu-E. Hu ◽  
...  
Keyword(s):  
Surface Area ◽  
High Surface ◽  
Enzymatic Catalysis ◽  
High Surface Area ◽  
Rhizomucor Miehei ◽  
Catalytic Performance ◽  
Biodiesel Production ◽  
Core Shell ◽  
Water Medium ◽  
Practical Applications

Abstract Background Production of biodiesel from renewable sources such as inedible vegetable oils by enzymatic catalysis has been a hotspot but remains a challenge on the efficient use of an enzyme. COFs (Covalent Organic Frameworks) with large surface area and porosity can be applied as ideal support to avoid aggregation of lipase and methanol. However, the naturally low density limits its application. In this work, we reported a facile synthesis of core–shell magnetic COF composite (Fe3O4@COF-OMe) to immobilize RML (Rhizomucor miehei lipase), to achieve its utilization in biodiesel production. Result This strategy gives extrinsic magnetic property, and the magnetic COFs is much heavier and could disperse in water medium well, facilitating the attachment with the enzyme. The resultant biocomposite exhibited an excellent capacity of RML due to its high surface area and fast response to the external magnetic field, as well as good chemical stability. The core–shell magnetic COF-OMe structure not only achieved highly efficient immobilization and recovery processes but also maintained the activity of lipase to a great extent. RML@Fe3O4@COF-OMe performed well in practical applications, while free lipase did not. The biocomposite successfully achieved the production of biodiesel from Jatropha curcas Oil with a yield of about 70% in the optimized conditions. Conclusion Magnetic COFs (Fe3O4@COF-OMe) for RML immobilization greatly improved catalytic performance in template reaction and biodiesel preparation. The magneticity makes it easily recovered and separated from the system. This first successful attempt of COFs-based immobilized enzyme broadened the prospect of biodiesel production by COFs with some inspiration.

scholarly journals Improved methanol tolerance of Rhizomucor miehei lipase based on N‑glycosylation within the α-helix region and its application in biodiesel production

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Miao Tian ◽  
Lingmei Yang ◽  
Zhiyuan Wang ◽  
Pengmei Lv ◽  
Junying Fu ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Rational Design ◽  
Rhizomucor Miehei ◽  
Raw Material ◽  
Biodiesel Production ◽  
High Catalytic Activity ◽  
Single Chain ◽  
Methanol Tolerance ◽  
First Choice ◽  
Α Helix

Abstract Background Liquid lipases are widely used to convert oil into biodiesel. Methanol-resistant lipases with high catalytic activity are the first choice for practical production. Rhizomucor miehei lipase (RML) is a single-chain α/β-type protein that is widely used in biodiesel preparation. Improving the catalytic activity and methanol tolerance of RML is necessary to realise the industrial production of biodiesel. Results In this study, a semi-rational design method was used to optimise the catalytic activity and methanol tolerance of ProRML. After N-glycosylation modification of the α-helix of the mature peptide in ProRML, the resulting mutants N218, N93, N115, N260, and N183 increased enzyme activity by 66.81, 13.54, 10.33, 3.69, and 2.39 times than that of WT, respectively. The residual activities of N218 and N260 were 88.78% and 86.08% after incubation in 50% methanol for 2.5 h, respectively. In addition, the biodiesel yield of all mutants was improved when methanol was added once and reacted for 24 h with colza oil as the raw material. N260 and N218 increased the biodiesel yield from 9.49% to 88.75% and 90.46%, respectively. Conclusions These results indicate that optimising N-glycosylation modification in the α-helix structure is an effective strategy for improving the performance of ProRML. This study provides an effective approach to improve the design of the enzyme and the properties of lipase mutants, thereby rendering them suitable for industrial biomass conversion.

scholarly journals Magnetic COFs as Satisfied Support for Lipase Immobilization and Recovery to Effectively Achieve the Production of Biodiesel by Great Maintenance of Enzyme Activity

2021 ◽  
Author(s):  
Zi-Wen Zhou ◽  
Chun-Xian Cai ◽  
Xiu Xing ◽  
Jun Li ◽  
Zu-E Hu ◽  
...  
Keyword(s):  
Surface Area ◽  
High Surface ◽  
Enzymatic Catalysis ◽  
High Surface Area ◽  
Rhizomucor Miehei ◽  
Catalytic Performance ◽  
Biodiesel Production ◽  
Core Shell ◽  
Water Medium ◽  
Practical Applications

Abstract Background: Production of biodiesel from renewable sources such as inedible vegetable oils by enzymatic catalysis has been a hotspot but remains a challenge on the efficient use of an enzyme. COFs (Covalent Organic Frameworks) with large surface area and porosity can be applied as ideal support to avoid aggregation of lipase and methanol. However, the naturally low density limits its application. In this work, we reported a facile synthesis of core-shell magnetic COF composite (Fe3O4@COF-OMe) to immobilize RML (Rhizomucor miehei lipase), to achieve its utilization in biodiesel production.Result: This strategy gives extrinsic magnetic property, and the magnetic COFs is much heavier and could disperse in water medium well, facilitating the attachment with the enzyme. The resultant biocomposite exhibited an excellent capacity of RML due to its high surface area and fast response to the external magnetic field, as well as good chemical stability. The core-shell magnetic COF-OMe structure not only achieved highly efficient immobilization and recovery processes but also maintained the activity of lipase to a great extent. RML@Fe3O4@COF-OMe performed well in practical applications, while free lipase did not. The biocomposite successfully achieved the production of biodiesel from Jatropha curcas Oil with a yield of about 70% in the optimized conditions. Conclusion: Magnetic COFs (Fe3O4@COF-OMe) for RML immobilization greatly improved catalytic performance in template reaction and biodiesel preparation. The magneticity makes it easily recovered and separated from the system. This first successful attempt of COFs-based immobilized enzyme broadened the prospect of biodiesel production by COFs with some inspiration.

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