Submit to Molecules Review for Molecules Propose a Special Issue

Journal Browser

► Journal Browser

Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes

Special Issue Editors
Special Issue Information
Keywords
Published Papers

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Bioorganic Chemistry".

Deadline for manuscript submissions: closed (15 November 2023) | Viewed by 23933

Share This Special Issue

Special Issue Editors

Dr. Ana Paula Tavares

E-Mail Website
Guest Editor

CICECO – Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal
Interests: enzymatic biocatalysis; proteins; enzymes; biopharmaceuticals; ionic liquids; biomolecules purification; aqueous biphasic systems; fermentation processes; enzyme immobilization; nanomaterials
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Valéria de Carvalho Santos Ebinuma

E-Mail
Guest Editor

Department of Engineering of Bioprocesses and Biotechnology, School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara, SP 14800-903, Brazil
Interests: enzymes; biopharmaceuticals; colorants; fermentation process; downstream process; biotechnological process

Special Issue Information

Dear Colleagues,

Considering the diversity of applications that can be done by enzymes, including pharmaceutical industry, food industry, textile industry, biosensors, environmental bioremediation, lignocellulose modification and organic synthesis, enzymes have been the focus of several studies and it will be one of the most important biocatalysts in the near future.

Since enzymes can display higher substrate specificity, enantio- and regio- selectivity, and improved stability, several industries are currently adding enzymatic approaches to their processes for developing greener and more efficient technologies. Moreover, enzymes need mild conditions to perform their bioreactions while leading to higher yields. In parallel, many efforts to enhance enzyme performance by means that may improve enzyme activity and stability have been intensified by immobilization techniques. So, due to their wide-ranging applications, enzymes are looked upon as potentially able to replace the conventional chemical industrial processes in several industries.

This Special Issue aims to cover promising, recent, and novel research trends in the use of enzymes as biocatalysts for the production of a variety of pharmaceutical, food, textile products and other biotechnological applications. Interdisciplinary research on novel concepts and approaches is also welcome.

Dr. Ana Tavares
Prof. Dr. Valéria de Carvalho Santos Ebinuma
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Molecules is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

biocatalysis
biopharmaceuticals
enzyme production
enzyme immobilization
enzyme purification
enzyme catalysis
enzyme engineering
enzymatic processes

Published Papers (11 papers)

Download All Papers

Research

Jump to: Review

15 pages, 2519 KiB

Open AccessArticle

Crystal Structure of Allantoinase from Escherichia coli BL21: A Molecular Insight into a Role of the Active Site Loops in Catalysis

by Yen-Hua Huang, Po-Chun Yang, En-Shyh Lin, Ya-Yeh Ho, Wei-Feng Peng, Hsin-Pin Lu, Chien-Chih Huang and Cheng-Yang Huang

Molecules 2023, 28(2), 827; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules28020827 - 13 Jan 2023

Cited by 3 | Viewed by 1528

Abstract

Allantoinase (ALLase; EC 3.5.2.5) possesses a binuclear metal center in which two metal ions are bridged by a posttranslationally carbamylated lysine. ALLase acts as a key enzyme for the biogenesis and degradation of ureides by catalyzing the conversion of allantoin into allantoate. Biochemically, ALLase belongs to the cyclic amidohydrolase family, which also includes dihydropyrimidinase, dihydroorotase, hydantoinase (HYDase), and imidase. Previously, the crystal structure of ALLase from Escherichia coli K-12 (EcALLase-K12) was reported; however, the two active site loops crucial for substrate binding were not determined. This situation would limit further docking and protein engineering experiments. Here, we solved the crystal structure of E. coli BL21 ALLase (EcALLase-BL21) at a resolution of 2.07 Å (PDB ID 8HFD) to obtain more information for structural analyses. The structure has a classic TIM barrel fold. As compared with the previous work, the two missed active site loops in EcALLase-K12 were clearly determined in our structure of EcALLase-BL21. EcALLase-BL21 shared active site similarity with HYDase, an important biocatalyst for industrial production of semisynthetic penicillin and cephalosporins. Based on this structural comparison, we discussed the functional role of the two active site loops in EcALLase-BL21 to better understand the substrate/inhibitor binding mechanism for further biotechnological and pharmaceutical applications. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

15 pages, 5637 KiB

Open AccessFeature PaperArticle

Safe Sialidase Production by the Saprophyte Oerskovia paurometabola: Gene Sequence and Enzyme Purification

by Rumyana Eneva, Stephan Engibarov, Yana Gocheva, Simona Mitova, Alexander Arsov, Kaloyan Petrov, Radoslav Abrashev, Irina Lazarkevich and Penka Petrova

Molecules 2022, 27(24), 8922; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27248922 - 15 Dec 2022

Cited by 1 | Viewed by 1508

Abstract

Sialidase preparations are applied in structural and functional studies on sialoglycans, in the production of sialylated therapeutic proteins and synthetic substrates for use in biochemical research, etc. They are obtained mainly from pathogenic microorganisms; therefore, the demand for apathogenic producers of sialidase is of exceptional importance for the safe production of this enzyme. Here, we report for the first time the presence of a sialidase gene and enzyme in the saprophytic actinomycete Oerskovia paurometabola strain O129. An electrophoretically pure, glycosylated enzyme with a molecular weight of 70 kDa was obtained after a two-step chromatographic procedure using DEAE cellulose and Q-sepharose. The biochemical characterization showed that the enzyme is extracellular, inductive, and able to cleave α(2→3,6,8) linked sialic acids with preference for α(2→3) bonds. The enzyme production was strongly induced by glycomacropeptide (GMP) from milk whey, as well as by sialic acid. Investigation of the deduced amino acid sequence revealed that the protein molecule has the typical six-bladed β-propeller structure and contains all features of bacterial sialidases, i.e., an YRIP motif, five Asp-boxes, and the conserved amino acids in the active site. The presence of an unusual signal peptide of 40 amino acids was predicted. The sialidase-producing O. paurometabola O129 showed high and constant enzyme production. Together with its saprophytic nature, this makes it a reliable producer with high potential for industrial application. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

17 pages, 4502 KiB

Open AccessArticle

Post-Consumer Poly(ethylene terephthalate) (PET) Depolymerization by Yarrowia lipolytica: A Comparison between Hydrolysis Using Cell-Free Enzymatic Extracts and Microbial Submerged Cultivation

by Julio Cesar Soares Sales, Aline Machado de Castro, Bernardo Dias Ribeiro and Maria Alice Zarur Coelho

Molecules 2022, 27(21), 7502; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27217502 - 03 Nov 2022

Cited by 1 | Viewed by 1352

Abstract

Several microorganisms have been reported as capable of acting on poly(ethylene terephthalate) (PET) to some extent, such as Yarrowia lipolytica, which is a yeast known to produce various hydrolases of industrial interest. The present work aims to evaluate PET depolymerization by Y. lipolytica using two different strategies. In the first one, biocatalysts were produced during solid-state fermentation (SSF-YL), extracted and subsequently used for the hydrolysis of PET and bis(2-hydroxyethyl terephthalate) (BHET), a key intermediate in PET hydrolysis. Biocatalysts were able to act on BHET, yielding terephthalic acid (TPA) (131.31 µmol L⁻¹), and on PET, leading to a TPA concentration of 42.80 µmol L⁻¹ after 168 h. In the second strategy, PET depolymerization was evaluated during submerged cultivations of Y. lipolytica using four different culture media, and the use of YT medium ((w/v) yeast extract 1%, tryptone 2%) yielded the highest TPA concentration after 96 h (65.40 µmol L⁻¹). A final TPA concentration of 94.3 µmol L⁻¹ was obtained on a scale-up in benchtop bioreactors using YT medium. The conversion obtained in bioreactors was 121% higher than in systems with SSF-YL. The results of the present work suggest a relevant role of Y. lipolytica cells in the depolymerization process. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

15 pages, 1956 KiB

Open AccessArticle

Production, Biochemical Characterization, and Kinetic/Thermodynamic Study of Inulinase from Aspergillus terreus URM4658

by Rodrigo Lira de Oliveira, Suzana Pedroza da Silva, Attilio Converti and Tatiana Souza Porto

Molecules 2022, 27(19), 6418; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27196418 - 28 Sep 2022

Cited by 5 | Viewed by 1223

Abstract

Inulinases are enzymes involved in the hydrolysis of inulin, which can be used in the food industry to produce high-fructose syrups and fructo-oligosaccharides. For this purpose, different Aspergillus strains and substrates were tested for inulinase production by solid-state fermentation, among which Aspergillus terreus URM4658 grown on wheat bran showed the highest activity (15.08 U mL⁻¹). The inulinase produced by this strain exhibited optimum activity at 60 °C and pH 4.0. A detailed kinetic/thermodynamic study was performed on the inulin hydrolysis reaction and enzyme thermal inactivation. Inulinase was shown to have a high affinity for substrate evidenced by very-low Michaelis constant values (0.78–2.02 mM), which together with a low activation energy (19.59 kJ mol⁻¹), indicates good enzyme catalytic potential. Moreover, its long half-life (t_1/2 = 519.86 min) and very high D-value (1726.94 min) at 60 °C suggested great thermostability, which was confirmed by the thermodynamic parameters of its thermal denaturation, namely the activation energy of thermal denaturation (E*_d = 182.18 kJ mol⁻¹) and Gibbs free energy (106.18 ≤ ΔG*_d ≤ 111.56 kJ mol⁻¹). These results indicate that A. terreus URM4658 inulinase is a promising and efficient biocatalyst, which could be fruitfully exploited in long-term industrial applications. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Graphical abstract

10 pages, 2115 KiB

Open AccessArticle

Production and Purification of Pectinase from Bacillus subtilis 15A-B92 and Its Biotechnological Applications

by Yahya S. Alqahtani, Sunil S. More, Keerthana R., Ibrahim Ahmed Shaikh, Anusha K. J., Veena S. More, Francois N. Niyonzima, Uday M. Muddapur and Aejaz A. Khan

Molecules 2022, 27(13), 4195; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27134195 - 29 Jun 2022

Cited by 10 | Viewed by 3331

Abstract

Enzymes that degrade pectin are called pectinases. Pectinases of microbial origin are used in juice clarification as the process is cost-effective. This study screened a pectinase-producing bacterium isolated from soil and identified as Bacillus subtilis 15A B-92 based on the 16S rRNA molecular technique. The purified pectinase from the isolate showed 99.6 U/mg specific activity and 11.6-fold purity. The molecular weight of the purified bacterial pectinase was 14.41 ± 1 kD. Optimum pectinase activity was found at pH 4.5 and 50 °C, and the enzyme was 100% stable for 3.5 h in these conditions. No enzymatic inhibition or activation effect was seen with Fe²⁺, Ca²⁺, or Mg²⁺. However, a slight inhibition was seen with Cu²⁺, Mn²⁺, and Zn²⁺. Tween 20 and 80 slightly inhibited the pectinase, whereas iodoacetic acid (IAA), ethylenediaminetetraacetate (EDTA), urea, and sodium dodecyl sulfate (SDS) showed potent inhibition. The bacterial pectinase degraded citrus pectin (100%); however, it was inactive in the presence of galactose. With citrus pectin as the substrate, the Km and Vmax were calculated as 1.72 mg/mL and 1609 U/g, respectively. The high affinity of pectinase for its substrate makes the process cost-effective when utilized in food industries. The obtained pectinase was able to clarify orange and apple juices, justifying its application in the food industry. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

9 pages, 1203 KiB

Open AccessArticle

Enzymatic Synthesis of Novel and Highly Soluble Puerarin Glucoside by Deinococcus geothermalis Amylosucrase

by Hsiou-Yu Ding, Tzi-Yuan Wang, Jiumn-Yih Wu, Yu-Li Tsai and Te-Sheng Chang

Molecules 2022, 27(13), 4074; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27134074 - 24 Jun 2022

Cited by 6 | Viewed by 1377

Abstract

Puerarin (daidzein-8-C-glucoside) is an isoflavone isolated from several leguminous plants of the genus Pueraria. Puerarin possesses several pharmacological properties; however, the poor solubility of puerarin limits its applications. To resolve this poor solubility, Deinococcus geothermalis amylosucrase (DgAS) was used to modify puerarin into more soluble derivatives. The results showed that DgAS could biotransform puerarin into a novel compound: puerarin-4′-O-α-glucoside. The biotransformation reaction was manipulated at different temperatures, pH values, sucrose concentrations, reaction times, and enzyme concentrations. The results showed that the optimal reaction condition was biotransformed by 200 μg/mL DgAS with 20% (w/v) sucrose at pH 6 and incubated at 40 °C for 48 h, and the optimal production yield was 35.1%. Puerarin-4′-O-α-glucoside showed 129-fold higher solubility than that of puerarin and, thus, could be further applied for pharmacological use in the future. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

16 pages, 2906 KiB

Open AccessArticle

Bioactive Properties of a Novel Antibacterial Dye Obtained from Laccase-Mediated Oxidation of 8-Anilino-1-naphthalenesulfonic Acid

by Jolanta Polak, Marcin Grąz, Kamila Wlizło, Katarzyna Szałapata, Justyna Kapral-Piotrowska, Roman Paduch and Anna Jarosz-Wilkołazka

Molecules 2022, 27(2), 487; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27020487 - 13 Jan 2022

Cited by 5 | Viewed by 1793

Abstract

Fungal laccase obtained from a Cerrena unicolor strain was used as an effective biocatalyst for the transformation of 8-anilino-1-naphthalenesulfonic acid into a green-coloured antibacterial compound, which can be considered as both an antimicrobial agent and a textile dye, simultaneously. The process of biosynthesis was performed in buffered solutions containing methanol as a co-solvent, allowing better solubilisation of substrate. The transformation process was optimised in terms of the buffer pH value, laccase activity, and concentrations of the substrate and co-solvent. The crude product obtained exhibited low cytotoxicity, antibacterial properties against Staphylococcus aureus and Staphylococcus epidermidis, and antioxidant properties. Moreover, the synthesised green-coloured compound proved non-allergenic and demonstrated a high efficiency of dyeing wool fibres. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

13 pages, 2330 KiB

Open AccessArticle

Engineering of Yeast Old Yellow Enzyme OYE3 Enables Its Capability Discriminating of (E)-Citral and (Z)-Citral

by Tairan Wang, Ran Wei, Yingting Feng, Lijun Jin, Yunpeng Jia, Duxia Yang, Zuonan Liang, Mengge Han, Xia Li, Chenze Lu and Xiangxian Ying

Molecules 2021, 26(16), 5040; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules26165040 - 20 Aug 2021

Cited by 6 | Viewed by 2208

Abstract

The importance of yeast old yellow enzymes is increasingly recognized for direct asymmetric reduction of (E/Z)-citral to (R)-citronellal. As one of the most performing old yellow enzymes, the enzyme OYE3 from Saccharomyces cerevisiae S288C exhibited complementary enantioselectivity for the reduction of (E)-citral and (Z)-citral, resulting in lower e.e. value of (R)-citronellal in the reduction of (E/Z)-citral. To develop a novel approach for the direct synthesis of enantio-pure (R)-citronellal from the reduction of (E/Z)-citral, the enzyme OYE3 was firstly modified by semi-rational design to improve its (R)-enantioselectivity. The OYE3 variants W116A and S296F showed strict (R)-enantioselectivity in the reduction of (E)-citral, and significantly reversed the (S)-enantioselectivity in the reduction of (Z)-citral. Next, the double substitution of OYE3 led to the unique variant S296F/W116G, which exhibited strict (R)-enantioselectivity in the reduction of (E)-citral and (E/Z)-citral, but was not active on (Z)-citral. Relying on its capability discriminating (E)-citral and (Z)-citral, a new cascade reaction catalyzed by the OYE3 variant S296F/W116G and glucose dehydrogenase was developed, providing the enantio-pure (R)-citronellal and the retained (Z)-citral after complete reduction of (E)-citral. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Figure 1

Review

Jump to: Research

52 pages, 10296 KiB

Open AccessReview

Direct Biocatalytic Processes for CO₂ Capture as a Green Tool to Produce Value-Added Chemicals

by Rocio Villa, Susana Nieto, Antonio Donaire and Pedro Lozano

Molecules 2023, 28(14), 5520; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules28145520 - 19 Jul 2023

Cited by 4 | Viewed by 2976

Abstract

Direct biocatalytic processes for CO₂ capture and transformation in value-added chemicals may be considered a useful tool for reducing the concentration of this greenhouse gas in the atmosphere. Among the other enzymes, carbonic anhydrase (CA) and formate dehydrogenase (FDH) are two key biocatalysts suitable for this challenge, facilitating the uptake of carbon dioxide from the atmosphere in complementary ways. Carbonic anhydrases accelerate CO₂ uptake by promoting its solubility in water in the form of hydrogen carbonate as the first step in converting the gas into a species widely used in carbon capture storage and its utilization processes (CCSU), particularly in carbonation and mineralization methods. On the other hand, formate dehydrogenases represent the biocatalytic machinery evolved by certain organisms to convert CO₂ into enriched, reduced, and easily transportable hydrogen species, such as formic acid, via enzymatic cascade systems that obtain energy from chemical species, electrochemical sources, or light. Formic acid is the basis for fixing C₁-carbon species to other, more reduced molecules. In this review, the state-of-the-art of both methods of CO₂ uptake is assessed, highlighting the biotechnological approaches that have been developed using both enzymes. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Graphical abstract

15 pages, 1303 KiB

Open AccessReview

Surfactants, Biosurfactants, and Non-Catalytic Proteins as Key Molecules to Enhance Enzymatic Hydrolysis of Lignocellulosic Biomass

by Salvador Sánchez-Muñoz, Thércia R. Balbino, Fernanda de Oliveira, Thiago M. Rocha, Fernanda G. Barbosa, Martha I. Vélez-Mercado, Paulo R. F. Marcelino, Felipe A. F. Antunes, Elisangela J. C. Moraes, Julio C. dos Santos and Silvio S. da Silva

Molecules 2022, 27(23), 8180; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27238180 - 24 Nov 2022

Cited by 10 | Viewed by 1999

Abstract

Lignocellulosic biomass (LCB) has remained a latent alternative resource to be the main substitute for oil and its derivatives in a biorefinery concept. However, its complex structure and the underdeveloped technologies for its large-scale processing keep it in a state of constant study trying to establish a consolidated process. In intensive processes, enzymes have been shown to be important molecules for the fractionation and conversion of LCB into biofuels and high-value-added molecules. However, operational challenges must be overcome before enzyme technology can be the main resource for obtaining second-generation sugars. The use of additives is shown to be a suitable strategy to improve the saccharification process. This review describes the mechanisms, roles, and effects of using additives, such as surfactants, biosurfactants, and non-catalytic proteins, separately and integrated into the enzymatic hydrolysis process of lignocellulosic biomass. In doing so, it provides a technical background in which operational biomass processing hurdles such as solids and enzymatic loadings, pretreatment burdens, and the unproductive adsorption phenomenon can be addressed. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures

Graphical abstract

17 pages, 980 KiB

Open AccessReview

Cold-Active Enzymes and Their Potential Industrial Applications—A Review

by Burhan Hamid, Zaffar Bashir, Ali Mohd Yatoo, Fayaz Mohiddin, Neesa Majeed, Monika Bansal, Peter Poczai, Waleed Hassan Almalki, R. Z. Sayyed, Ali A. Shati and Mohammad Y. Alfaifi

Molecules 2022, 27(18), 5885; https://0-doi-org.brum.beds.ac.uk/10.3390/molecules27185885 - 10 Sep 2022

Cited by 18 | Viewed by 3245

Abstract

More than 70% of our planet is covered by extremely cold environments, nourishing a broad diversity of microbial life. Temperature is the most significant parameter that plays a key role in the distribution of microorganisms on our planet. Psychrophilic microorganisms are the most prominent inhabitants of the cold ecosystems, and they possess potential cold-active enzymes with diverse uses in the research and commercial sectors. Psychrophiles are modified to nurture, replicate, and retain their active metabolic activities in low temperatures. Their enzymes possess characteristics of maximal activity at low to adequate temperatures; this feature makes them more appealing and attractive in biotechnology. The high enzymatic activity of psychrozymes at low temperatures implies an important feature for energy saving. These enzymes have proven more advantageous than their mesophilic and thermophilic counterparts. Therefore, it is very important to explore the efficiency and utility of different psychrozymes in food processing, pharmaceuticals, brewing, bioremediation, and molecular biology. In this review, we focused on the properties of cold-active enzymes and their diverse uses in different industries and research areas. This review will provide insight into the areas and characteristics to be improved in cold-active enzymes so that potential and desired enzymes can be made available for commercial purposes. Full article

(This article belongs to the Special Issue Biocatalysis and Enzymes in Pharmaceutical and Biotechnological Processes)

► Show Figures