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Advances and Applications in Molecular Enzymology

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".

Deadline for manuscript submissions: 20 May 2025 | Viewed by 875

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Guest Editor
Institute of Science and Technology-UFVJM, Diamantina, Minas Gerais, Brazil
Interests: food science and biotechnology; industrial biotechnology; bioconersion processes; bioactive potential of metabolites and ingredients
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Special Issue Information

Dear Colleagues,

Microbes are the key for the production of various valuable enzymes and the application of these enzymes to bioproduct development. Microbial enzymes offer extensive and increasingly significant advantages over chemical catalysts since they are derived from renewable resources and are biodegradable. Molecular enzyme technology has become one of the vital research areas in the field of natural sciences. Over five hundred industrial enzymes are being used for many applications in the agri-food-pharma-environment and other related sectors and constitute a multibillion USD annual market globally. From the molecular enzymology perspective, microbial bioengineering and applied biological sciences approaches, like metabolic engineering, next-generation sequencing, functional genomics, systems, and synthetic biology are the key to engineering microbial cell factories to produce tailor-made enzyme bioprocess. The growing demand for tailor-made enzymes from microorganisms for various applications requires the development of new biosynthetic and bioengineering tools, produced from fungi and bacterial systems. Microbial enzyme engineering includes biotechnological innovation, chemical engineering, drug developments, and applied biorefineries development systems. The application of nanotechnology in enzyme bioprocess design and production has opened new avenues for the application of enzymes in the various sectors, especially in biopharmaceutical and drug delivery systems, helping in designing new and novel recombinant proteins and glycans.

This Special Issue also welcomes submissions exploiting diverse and novel microbes from various extremes/hotspots as pillars for the industry and academic research to develop microbial molecular enzymology platform with potential applications in biopharmaceuticals, biochemicals, and several other relevant sectors.

This Special Issue will publish articles including reviews and research papers with an emphasis on enzyme production and formulations.

Prof. Dr. Gustavo Molina
Guest Editor

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Keywords

  • microbial enzymes
  • biosynthesis and bioengineering
  • drug development and targets
  • enzymes in the environment
  • engineered enzymes
  • enzymes in agri-food
  • enzymatic formulations
  • enzyme engineering
  • enzymes in biomedical sciences
  • green chemistry/technology
  • glycans
  • molecular evolution
  • nanocatalyst and nanotechnology
  • pharma enzymes
  • recombinant proteins production and purification
  • system and synthetic biology

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Published Papers (1 paper)

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Research

19 pages, 12976 KiB  
Article
Construction of Tandem Multimers with Different Combinatorial Forms of BmSPI38 and BmSPI39 and Analysis of Their Expression and Activity in Escherichia coli
by Zhaofeng Zhang, Youshan Li, Xi Yang, Changqing Chen, Shuai Ru, Jie Jiang, Wenyao Cai, Jiyu Li, Juanle Du and Dejue Qiao
Int. J. Mol. Sci. 2025, 26(5), 1788; https://doi.org/10.3390/ijms26051788 - 20 Feb 2025
Viewed by 422
Abstract
It was found that the serine protease inhibitors BmSPI38 and BmSPI39 in silkworm can strongly inhibit the activity of porcine pancreatic elastase, which has potential applicational value in the drug research and development of lung diseases, inflammatory diseases, and skin aging caused by [...] Read more.
It was found that the serine protease inhibitors BmSPI38 and BmSPI39 in silkworm can strongly inhibit the activity of porcine pancreatic elastase, which has potential applicational value in the drug research and development of lung diseases, inflammatory diseases, and skin aging caused by the excessive release of elastase. Previous studies have shown that homotypic multimers obtained by tandem expression can significantly enhance the antifungal activity and structural homogeneity of BmSPI38 and BmSPI39, while the effect of the tandem expression of these two inhibitors, with different combinations, on the total activity and expression levels of multimers remains unclear. The aim of this study is to explore whether it is possible to obtain the combination of BmSPI38 and BmSPI39 with strong total expression activity by protein engineering. In this study, 40 tandem multimer expression vectors with different combinatorial forms of BmSPI38 and BmSPI39 were constructed by the isocaudomer method, and recombinant proteins were obtained by the prokaryotic expression system. The target proteins were separated by SDS-PAGE to analyze the expression levels of multimer proteins with different combinatorial forms. The total activity of the recombinant expression products with different tandem forms was investigated using the in-gel activity staining technique of protease inhibitors. The SDS-PAGE results show that the expression levels of tandem multimers containing the BmSPI39 module at the carboxyl terminus were generally higher in the Escherichia coli supernatant than that of the tandem multimers containing the BmSPI38 module at the carboxyl terminus. The activity staining results indicate that compared with BmSPI38 and BmSPI39 homotypic multimers, the total activity of some recombinant expression products with different tandem forms was stronger. Furthermore, the total activity level was relatively higher when the carboxyl terminus of the multimer was a BmSPI39 module, such as the tandem dimers SPIAB and SPIaB and the tandem trimers SPIabB, SPIaaB, and SPIbaB. In this study, the expression of tandem fusion proteins with different combinations of the silkworm protease inhibitors BmSPI38 and BmSPI39 in E. coli was successfully achieved. It was confirmed that the tandem of different combinatorial forms, based on protein engineering, was an effective way to enhance the total activity of the fusion proteins of BmSPI38 and BmSPI39 and to improve their expression levels. Additionally, a number of multimer proteins with strong total activity and high exogenous expression levels were also screened, for example, SPIbaA, SPIbbA, SPIbbB, SPIabB, SPIaaB, and SPIbaB. This study not only lays the foundation for the exogenous production and development of BmSPI38 and BmSPI39 but also provides a reference for the construction of tandem and multimerization exploration of other protease inhibitors. Full article
(This article belongs to the Special Issue Advances and Applications in Molecular Enzymology)
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