Article
Biotechnology & Applied Microbiology
Fidan Erden-Karaoglan, Mert Karaoglan
Summary: This study investigated the applicability of heterologous yeast promoters in Pichia pastoris, and found inducible and constitutive regulation modes as well as the optimal carbon sources for these promoters. The study also determined the DNA region with the highest activity for all promoters through promoter deletion analysis.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Wenyuan Zhou, Sheng Tong, Farrukh Raza Amin, Wuxi Chen, Jinling Cai, Demao Li
Summary: In this study, endoglucanase (MtEG5-1) from Myceliophthora thermophila was successfully overexpressed in Pichia pastoris GS115. The purified MtEG5-1 showed optimal activity at pH 5 and 70 degrees C, with good thermal stability. The enzyme demonstrated a molecular weight of 55 kDa and had Km and Vmax values of approximately 6.11 mg/mL and 91.74 mu mol/min/mg at 70 degrees C (pH 5.0), respectively. The activity of MtEG5-1 was enhanced by Mn2+ and inhibited by K+. These findings suggest that MtEG5-1 expressed by P. pastoris GS115 is more heat-tolerant and suitable for future academic research and industrial applications.
FERMENTATION-BASEL
(2023)
Article
Biotechnology & Applied Microbiology
Yafei Zhang, Shengyan Wang, Lingfang Lu, Chenshan Zhang, Fan Cai, Yao Lin, Yide Huang
Summary: This study identified 18 promoters with high transcription levels under different carbon sources, with 9 of them showing higher protein expression compared to pGAP. The methanol-induced pFDH1 exhibited slightly higher protein expression than commonly used pAOX1, and was not repressed by glycerol.
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Nai-Xin Lin, Rui-Zhen He, Yan Xu, Xiao-Wei Yu
Summary: The robust P. pastoris mutant demonstrates higher expression levels of not only lipase but also other heterogeneous proteins under methanol induction conditions. The mutant shows lower intracellular ROS levels compared to the WT. RNA-seq analysis revealed upregulation of key pathways related to antioxidant, MAPK pathway, ergosterol synthesis pathway, transcription factors, and the peroxisome pathway in the robust mutant, contributing to its enhanced oxidative stress tolerance and high heterologous protein expression efficiency.
BIOTECHNOLOGY FOR BIOFUELS
(2021)
Article
Biochemistry & Molecular Biology
Samin Haghighi Poodeh, Seyed Omid Ranaei Siadat, Sareh Arjmand, Maryam Khalifeh Soltani
Summary: This study presents a novel Pichia pastoris strain engineered to co-express methanol-induced transcription factor 1 (Mit1) and the target protein. The overexpression of Mit1 upregulates the expression of the target recombinant proteins and enhances the yeast cells' capacity for methanol detoxification.
MOLECULAR BIOLOGY REPORTS
(2022)
Article
Chemistry, Multidisciplinary
Roberto Carlos Aristicas Ribalta, Lisandra Martinez Valdes, Meinardo Lafargue Gamez, Sonia Rodriguez Davydenko, Eric Dubreucq, Veronique Perrier, Benoit Moreau, Reinaldo Fraga Vidal
Summary: This study reveals the potential application of a novel dextranase variant in the sugar industry, which demonstrates advantages in both production yield and efficiency. It also provides evidence of carbohydrate-protein interactions.
APPLIED SCIENCES-BASEL
(2022)
Article
Biochemical Research Methods
Pascal Puellmann, Martin J. Weissenborn
Summary: Fungal peroxygenases (UPOs) have attracted significant interest as oxyfunctionalization catalysts, but their widespread use in biocatalysis is limited by challenges in heterologous production. This study developed a promoter and signal peptide shuffling system for high throughput UPO production in yeast, successfully producing new UPOs and demonstrating feasibility for other enzyme classes. The system achieved high production titers and promising yields without methanol, showcasing its potential for industrial applications.
ACS SYNTHETIC BIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Adivitiya Babbal, Shilpa Mohanty, Yogender Pal Khasa
Summary: A tag-free variant of streptokinase with improved production and stability was developed using a modified fermentation process and a combined feeding strategy, resulting in a significant increase in yield and protein stability.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Biotechnology & Applied Microbiology
Na Zhan, Licong Zhang, Hong Yang, Yalan Zheng, Xinke Wei, Jiajun Wang, Anshan Shan
Summary: This study improved the antimicrobial activity of LL37 by amino acid substitution and dimeric design, successfully expressing recombinant LG with strong antimicrobial activity and low hemolytic activity, as well as improved tolerance to salt ions compared to monomeric peptide FR.
MICROBIAL CELL FACTORIES
(2021)
Article
Biochemical Research Methods
Zhiqian Wang, Yunjun Yan, Houjin Zhang
Summary: Pichia pastoris has traditionally been regulated through methanol induction, which is toxic and flammable. In this study, a novel one-component optogenetic system based on the photosensitive protein EL222 was designed to induce gene expression under blue light and rapidly deactivate it upon switching back to darkness. The intensity of induction in the optogenetic system correlates positively with the dose and periodicity of blue light, and it demonstrates good spatial control.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Immunology
Nicholas J. MacDonald, Kavita Singh, Karine Reiter, Nguyen Vu, Richard Shimp, Apostolos G. Gittis, Beth Chen, Martin Burkhardt, Baoshan Zhang, Zhixiong Wang, Raul Herrera, Mackenzie Moler, Duck-Yeon Lee, Sachy Orr-Gonzalez, Jessica Herrod, Lynn E. Lambert, Kelly M. Rausch, Olga Muratova, David S. Jones, Yimin Wu, Albert J. Jin, David N. Garboczi, Patrick E. Duffy, David L. Narum
Summary: The development of a malaria vaccine that can block transmission of the parasite from mosquitoes to humans is crucial for malaria elimination efforts. The use of Pfs25, a protein on the surface of the parasite, is being investigated as a transmission-blocking vaccine. Two recombinant forms of Pfs25, Pfs25H and Pfs25M, have been studied for their structural and immunological differences.
Article
Microbiology
Andrey Rumyantsev, Anton Sidorin, Artemii Volkov, Ousama Al Shanaa, Elena Sambuk, Marina Padkina
Summary: This study analyzed the complex effects of proline on gene expression in Komagataella phaffii yeast, revealing differential expression of around 18.9% of protein-encoding genes in proline-containing media. The presence of proline does not inhibit the catabolism of proline by K. phaffii, and it represses genes involved in the utilization of other element sources. The repression of the AOX1 gene promoter with proline can be partially reversed by the deletion of the KpPUT4.2 gene.
Article
Biotechnology & Applied Microbiology
Ana Lucia Herrera-Estala, Jose Antonio Fuentes-Garibay, Martha Guerrero-Olazaran, Jose Maria Viader-Salvado
Summary: In this study, we constructed a constitutive P. pastoris expression system to extracellularly produce the phytase FTEII. Optimization of the culture conditions led to increased phytase productivity, and the results showed that the regulation of specific growth rate and temperature played a key role in enhancing phytase production, rather than increasing heterologous gene transcription.
JOURNAL OF BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Duniesky Martinez, Carmen Menendez, Osmani Chacon, Alejandro D. Fuentes, Dalia Borges, Alina Sobrino, Ricardo Ramirez, Enrique R. Perez, Lazaro Hernandez
Summary: The gene construct encoding Talaromyces minioluteus dextranase (r-TmDEX) was expressed in Pichia pastoris with an increase in transgene dosage resulting in higher extracellular yield of the enzyme. The optimal activity of the N-glycosylated r-TmDEX was observed at pH 4.5-5.5 and temperature 50-60 degrees C, while the addition of sucrose as a stabilizer maintained enzyme activity under harsh conditions. The use of the P. pastoris pGAP-based expression system proved to be suitable for large-scale production of dextranase and its application in dextran removal.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Ahmad Raza, Saira Bashir, Ratnasri Pothula, Heba Abdelgaffar, Romana Tabassum, Muhammad Irfan Anwar, Mian Muhammad Awais, Masood Akhtar, Juan Luis Jurat-Fuentes
Summary: Exogenous enzymes such as BsEgl and Bteq beta gluc added to poultry feed can improve the nutritional value of feed, enhance growth performance, and optimize feed conversion ratio. The study results suggest that supplementing high-fiber diets with recombinant enzymes can effectively improve the nutritional performance of poultry, particularly commercial broilers.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Review
Biotechnology & Applied Microbiology
Yasser Gaber, Boshra Rashad, Rasha Hussein, Mai Abdelgawad, Nourhan S. Ali, Tarek Dishisha, Aniko Varnai
BIOTECHNOLOGY ADVANCES
(2020)
Article
Multidisciplinary Sciences
Antonielle Monclaro, Dejan M. Petrovic, Gabriel S. C. Alves, Marcos M. C. Costa, Glaucia E. O. Midorikawa, Robert N. G. Miller, Edivaldo X. F. Filho, Vincent G. H. Eijsink, Aniko Varnai
Review
Biochemistry & Molecular Biology
Yuka Kojima, Aniko Varnai, Vincent G. H. Eijsink, Makoto Yoshida
TRENDS IN GLYCOSCIENCE AND GLYCOTECHNOLOGY
(2020)
Review
Biotechnology & Applied Microbiology
Heidi Ostby, Line Degn Hansen, Svein J. Horn, Vincent G. H. Eijsink, Aniko Varnai
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
(2020)
Article
Biotechnology & Applied Microbiology
Adnan Kadic, Aniko Varnai, Vincent G. H. Eijsink, Svein Jarle Horn, Gunnar Liden
Summary: Biochemical conversion of lignocellulosic biomass to sugars is hindered by high saccharifying enzyme costs. LPMOs offer potential solutions, but overdosing H2O2 can lead to enzyme inactivation. In this study, real-time monitoring of oxidation-reduction potential and H2O2 concentration allowed for predicting LPMO inactivation during enzymatic saccharification, potentially improving overall yields.
BIOTECHNOLOGY FOR BIOFUELS
(2021)
Article
Biotechnology & Applied Microbiology
Monika Tolgo, Olav A. Hegnar, Heidi Ostby, Aniko Varnai, Francisco Vilaplana, Vincent G. H. Eijsink, Lisbeth Olsson
Summary: This study provides a detailed functional characterization of six AA9 LPMOs from Thermothielavioides terrestris LPH172. The research shows that these LPMOs have different activities on various cellulose substrates, further supporting the hypothesis that the multiplicity of LPMO genes is related to the complex structure of lignocellulosic biomass.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Olav A. Hegnar, Heidi Ostby, Dejan M. Petrovic, Lisbeth Olsson, Aniko Varnai, Vincent G. H. Eijsink
Summary: Certain AA9 LPMOs were found to preferentially cleave xylan when acting on a cellulose-glucuronoxylan mixture, and this ability is due to protein evolution resulting in specific structural features. These findings strengthen the idea that the vast arsenal of AA9 LPMOs in certain fungal species provides functional versatility and may have evolved to promote oxidative depolymerization of a wide variety of recalcitrant, copolymeric plant polysaccharide structures.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Fredrik Gjerstad Stopamo, Asmund Kjendseth Rohr, Sophanit Mekasha, Dejan M. Petrovic, Aniko Varnai, Vincent G. H. Eijsink
Summary: The study reports the structural and functional characteristics of a single-domain AA11 LPMO from Aspergillus fumigatus. The LPMO, AfAA11A, shows substrate-binding surface features similar to those of known chitin-active LPMOs and exhibits considerable affinity for both alpha-chitin and beta-chitin, enhancing chitin degradation.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2021)
Article
Chemistry, Multidisciplinary
Line Degn Hansen, Martin Ostensen, Bjornar Arstad, Roman Tschentscher, Vincent G. H. Eijsink, Svein J. Horn, Aniko Varnai
Summary: The use of a carbocation scavenger to improve the enzymatic saccharification of steam-exploded softwood brings us closer to a softwood-based biorefinery. The impregnation effect enables complete saccharification of spruce cellulose with lytic polysaccharide monooxygenase (LPMO)-containing Cellic CTec2, but not with an LPMO-poor cellulase cocktail (Celluclast).
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2022)
Article
Plant Sciences
Andre van Eerde, Aniko Varnai, Yanliang Wang, Lisa Paruch, John-Kristian Jameson, Fen Qiao, Hans Geir Eiken, Hang Su, Vincent G. H. Eijsink, Jihong Liu Clarke
Summary: Giant pandas have been able to exclusively consume bamboo as their diet for 2 million years due to the enzymes produced by their gut bacteria. A bacterial laccase, Lac51, was identified in the gut microbiome of giant pandas and expressed in non-food plants, showing the potential of using green plants as a platform for bacterial laccase production.
FRONTIERS IN PLANT SCIENCE
(2022)
Article
Chemistry, Multidisciplinary
Heidi Ostby, Aniko Varnai, Raphael Gabriel, Piotr Chylenski, Svein J. Horn, Steven W. Singer, Vincent G. H. Eijsink
Summary: Understanding and improving the efficiency of enzymatic saccharification of lignocellulosic biomass is crucial for utilizing renewable materials. This study focuses on the role of lytic polysaccharide monooxygenases (LPMOs) and investigates various process parameters. The results demonstrate the potential of LPMO-rich cellulolytic secretome and the usefulness of glucose oxidase for optimizing saccharification.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2022)
Article
Biotechnology & Applied Microbiology
Line D. Hansen, Vincent G. H. Eijsink, Svein J. Horn, Aniko Varnai
Summary: Simultaneous saccharification and fermentation (SSF) is a commonly used strategy for lignocellulosic biomass valorization. Adding small amounts of H2O2 during SSF can sustain the activity of lytic polysaccharide monooxygenases (LPMOs) and improve cellulose depolymerization efficiency, leading to increased lactic acid production.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Review
Biochemistry & Molecular Biology
Heidi ostby, Aniko Varnai
Summary: This paper reviews the potential of hemicellulose in lignocellulose processing, focusing on what can be achieved using enzymatic means. The various enzyme activities required for complete depolymerization of the primary hemicellulose types found in plant cell walls and the upgrading of hemicellulosic polymers, oligosaccharides, and pentose sugars derived from hemicellulose depolymerization into a broad spectrum of value-added products are discussed.
ESSAYS IN BIOCHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Tina R. Tuveng, Heidi Ostby, Ketty C. Tamburrini, Bastien Bissaro, Olav A. Hegnar, Anton A. Stepnov, Aniko Varnai, Jean-Guy Berrin, Vincent G. H. Eijsink
Summary: Lytic polysaccharide monooxygenases (LPMOs) belonging to the AA14 family are believed to contribute to the enzymatic degradation of lignocellulosic biomass by specifically acting on xylan in recalcitrant cellulose-xylan complexes. However, the functional characterization of two AA14 LPMOs, TrAA14A from Trichoderma reesei and PcoAA14A from Pycnoporus coccineus, showed that these enzymes have oxidase and peroxidase activities but their actual substrate remains unknown. The findings raise questions about the nature of AA14 LPMOs and highlight challenges in their functional characterization.
Review
Biochemistry & Molecular Biology
Yogesh B. Chaudhari, Aniko Varnai, Morten Sorlie, Svein J. Horn, Vincent G. H. Eijsink
Summary: Lignocellulosic biomass is a valuable renewable resource for energy, chemicals, and materials. Efficient enzymatic depolymerization of cellulose to glucose is essential for its economically viable utilization. This review focuses on the engineering of cellulases to improve their efficiency, stability, and cost-effectiveness. The review also provides an overview of recent advancements and notable studies in cellulase engineering.
PROTEIN ENGINEERING DESIGN & SELECTION
(2023)