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
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, Physical
Xiaoxu Yang, Yan Zhang, Huimin Pang, Sheng Yuan, Xuxia Wang, Zhiming Hu, Qinghua Zhou, Yaojia He, Yunjun Yan, Li Xu
Summary: The study successfully produced biodiesel from tallow seed oil by codisplaying CRL1 and ROL on the cell surfaces of P. pastoris whole-cell catalysts. The activity of codisplaying strains was significantly higher than that of single-displayed enzymes, and using codisplayed enzymes for catalysis can increase the production of methyl esters.
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
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
Microbiology
Tatiana Ianshina, Anton Sidorin, Kristina Petrova, Maria Shubert, Anastasiya Makeeva, Elena Sambuk, Anastasiya Govdi, Andrey Rumyantsev, Marina Padkina
Summary: The effect of methionine on gene expression in Komagataella phaffii yeast cells was investigated, revealing its impact on genes involved in methionine biosynthesis, fatty acid metabolism, and methanol utilization. This study provides important information for optimizing media recipes and cultivation strategies to maximize the efficiency of recombinant product synthesis.
Article
Biochemical Research Methods
Amuliya Kashyap, Kuldeep Saini, Meenu Saini, Yogender Pal Khasa, Rani Gupta
Summary: A novel Lip(+) Pichia pastoris expression platform was developed, integrating lipase Lip2 from Yarrowia lipolytica under the constitutive Glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. Effective expression of amylase from Bacillus licheniformis was achieved utilizing methyloleate in Lip(+)Amy(+) host. The use of methyloleate led to comparable protein expression and increased biomass production.
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Yanelis Robainas-del-Pino, Jose Maria Viader-Salvado, Ana Lucia Herrera-Estala, Martha Guerrero-Olazaran
Summary: This study identified and functionally characterized the promoter and transcriptional terminator of a constitutive gene, 1033, in the yeast Komagataella phaffii. The results showed that the promoter of 1033 had 2-3% transcriptional activity, which was tunable by cell growth and carbon source. Different combinations of regulatory elements affected the transcriptional activity of heterologous and endogenous genes, and were dependent on the carbon source. The promoter-terminator pair and carbon source influenced the translation and/or protein secretion pathway of the heterologous gene, with low transcript levels and glycerol cultures promoting translation and secretion.
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Qi Shen, Zhuang Yu, Xiao-Ting Zhou, Shi-Jia Zhang, Shu-Ping Zou, Neng Xiong, Ya-Ping Xue, Zhi-Qiang Liu, Yu-Guo Zheng
Summary: A novel glucose-regulated promoter, P-Dog2p300, was identified for driving the expression of antibiotic-resistant genes in Pichia pastoris, resulting in reduced metabolic burden during recombinant protein expression. The use of P-Dog2p300 effectively selected transformants harboring multiple integrations in a cost-effective manner.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(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
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
Biochemical Research Methods
Fidan Erden-Karaogian, Mert Karaogian, Gurkan Yilmaz, Semiramis Yilmaz, Mehmet Inan
Summary: This study investigated the regulation of the ADH genes in P. pastoris, identifying the UAS and URS sequences of the ADH2 promoter. Five synthetic promoters were constructed by altering regulatory sites, leading to improved xylanase production. The highest activity synthetic promoter, SNT5, was further evaluated at the fermenter scale, with potential implications for industrial processes.
BIOTECHNOLOGY JOURNAL
(2022)
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
Diego Noseda, Cecilia D'Alessio, Javier Santos, Tommy Idrovo-Hidalgo, Florencia Pignataro, Diana E. Wetzler, Hernan Gentili, Alejandro D. Nadra, Ernesto Roman, Carlos Pavan, Lucas A. M. Ruberto
Summary: SARS-CoV-2 is the pathogenic agent causing the COVID-19 pandemic. The Spike protein, an external and exposed protein encoded by this virus, interacts with the ACE2 receptors of human cells through a receptor binding domain (RBD), allowing virus entry. This study developed a method for producing recombinant RBD using Pichia pastoris in a stirred-tank bioreactor (SRTB) up to 7 L. By utilizing a four-stage procedure with a basic saline medium containing glycerol, methanol, and compressed air, approximately 500 mg/L of raw RBD and 206 mg/L of purified (>95%) RBD were successfully obtained. Therefore, this proposed method offers a feasible, simple, scalable, and cost-effective approach for obtaining RBD for diagnosis kits and vaccine formulation.
FERMENTATION-BASEL
(2023)
Article
Biochemistry & Molecular Biology
Dunchi Xiao, Xun Li, Yu Zhang, Fei Wang
Summary: In this study, different promoters and signal peptides were compared for expressing codon-optimized CALB in Pichia pastoris. Recombinant P. pastoris containing 3 copies of calb gene were obtained by screening with high concentrations of antibiotics. The maximum CALB activity and total protein content reached 444.46 +/- 18.81 U/mL and 5.41 +/- 0.1 g/L, respectively, after about 9 days of incubation in FM22 medium.
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
(2023)
Article
Biochemical Research Methods
Juan J. Barrero, Alejandro Pagazartaundua, Benjamin S. Glick, Francisco Valero, Pau Ferrer
Summary: Pichia pastoris has been identified as having a bottleneck at the ER translocation level, with an improved secretion signal leading to increased production of target proteins and supporting higher maximum specific growth rates.
Review
Biochemistry & Molecular Biology
Sonakshi De, Diethard Mattanovich, Pau Ferrer, Brigitte Gasser
Summary: Pichia pastoris has become the most popular yeast cell factory for producing heterologous proteins, with strong promoters, stable genetic constructs, and a growing collection of strains, tools, and protocols. In addition to small-scale production for biochemical studies and protein characterization, the yeast is also being increasingly used for in vivo biotransformation and metabolic pathway engineering for chemical production.
MICROBIAL CELL FACTORIES-BOOK
(2021)
Article
Agricultural Engineering
Josu Lopez-Fernandez, Maria Dolors Benaiges, Francisco Valero
Summary: Immobilizing Rhizopus oryzae lipase on Purolite (R) and magnetite superparamagnetic supports increased operational stability; functionalizing Purolite (R) enhanced biodiesel and volumetric productivity; raising substrate acidity significantly improved operational stability of the biocatalyst.
BIORESOURCE TECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Albert Fina, Gabriela Coelho Breda, Miriam Perez-Trujillo, Denise Maria Guimaraes Freire, Rodrigo Volcan Almeida, Joan Albiol, Pau Ferrer
Summary: This study demonstrated the use of P. pastoris as a cell factory to produce 3-hydroxypropionic acid from glycerol. By introducing the mcr gene from Chloroflexus aurantiacus and optimizing enzyme and metabolic engineering modifications, a significant increase in 3-HP production was achieved. The best strain (PpHP6) showed a high volumetric productivity, indicating the promising potential of P. pastoris for the production of bulk chemicals, especially 3-HP, from crude glycerol.
MICROBIAL BIOTECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Manuel Alarcon-Vivero, Nathaly Ruiz-Tagle Moena, Fidelina Gonzalez, Paz Jopia-Contreras, Estrella Aspe, Homero Urrutia Briones, Katherine Sossa Fernandez
Summary: The inhibitory effect of ammonia on methane production can be controlled by using acclimated biofilms enriched with ammonia tolerant methanogens.
BIOTECHNOLOGY LETTERS
(2022)
Article
Chemistry, Physical
Josu Lopez-Fernandez, Maria Dolors Benaiges, Francisco Valero
Summary: In this study, the Rhizopus oryzae lipase (ROL) with a 27-amino acid prosequence was successfully expressed in Komagataella phaffii, resulting in significantly higher activity and productivity compared to methanol-induced expression. The truncated prosequence enabled more efficient production of proROL and the immobilized form showed increased stability in biodiesel and ethyl butyrate production processes.
Article
Energy & Fuels
Josu Lopez-Fernandez, Desiree Moya, Maria Dolors Benaiges, Francisco Valero, Manel Alcala
Summary: This study demonstrates the use of NIR spectroscopy for inline monitoring of enzymatic production of biodiesel from waste cooking oil. The results show a high correlation between NIR spectroscopy and gas chromatography, indicating that NIR spectroscopy is suitable for real-time monitoring of enzymatic production of biodiesel.
Article
Biotechnology & Applied Microbiology
Martina Pasini, Alfred Fernandez-Castane, Gloria Caminal, Tim W. Overton, Pau Ferrer
Summary: This study investigates the suitability of a set of IPTG-inducible protein expression systems in bioprocess conditions resembling industrial settings, and successfully improves the production yield and advantageous features of recombinant proteins through several generations of engineering.
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
(2022)
Article
Chemistry, Physical
Josu Lopez-Fernandez, Maria Dolors Benaiges, Xavier Sebastian, Jose Maria Bueno, Francisco Valero
Summary: Enzymatic synthesis of short-chain esters using covalently immobilised Rhizopus oryzae lipase was investigated. The best yield of isoamyl butyrate (ca. 100%) was obtained in cyclohexane as the reaction solvent. Optimum initial acid concentration and acid:alcohol mole ratio were determined to maximize the production of isoamyl butyrate with reused enzyme. The biocatalyst and reaction conditions proposed in this study could be used for industrial production of isoamyl esters.
Article
Biotechnology & Applied Microbiology
Albert Fina, Stephanie Heux, Joan Albiol, Pau Ferrer
Summary: This study improves the productivity of Pichia pastoris in producing 3-hydroxypropionic acid by combining different metabolic engineering approaches. The results show that, in combination with using glycerol as a carbon source, the metabolically engineered strains achieved the highest concentration and productivity of 3-hydroxypropionic acid reported so far in yeast.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Review
Chemistry, Physical
Ana Laura Reyes-Reyes, Francisco Valero Barranco, Georgina Sandoval
Summary: This review discusses the potential applications of lipases in the nutraceutical and food industry, focusing on the production of high-value compounds and recent successful cases of modified compounds produced using lipase reactions.
Correction
Biotechnology & Applied Microbiology
Albert Fina, Stephanie Heux, Joan Albiol, Pau Ferrer
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Albert Fina, Pierre Millard, Joan Albiol, Pau Ferrer, Stephanie Heux
Summary: By using a high throughput C-13-metabolic flux analysis platform, the metabolic phenotypes of 10 3-HP producing P. pastoris strains were characterized, revealing the impact of NADPH regeneration and cytosolic acetyl-CoA synthesis pathway on metabolic flux distribution. Additionally, the metabolic adaptation of P. pastoris to acidic pH was also investigated.
MICROBIAL CELL FACTORIES
(2023)
Article
Biotechnology & Applied Microbiology
Gabriela C. Breda, Priscila E. Faria, Yuri S. Rodrigues, Priscila B. Pinheiro, Maria Clara R. Nucci, Pau Ferrer, Denise M. G. Freire, Rodrigo V. Almeida, Rafael D. Mesquita
Summary: ProspectBIO is a new software for bioprospecting that can efficiently find non-annotated enzyme sequences by searching for homologs in genomes. Using this software, a novel lipase-encoding gene from Ustilago hordei was discovered and successfully expressed in yeast, resulting in an active enzyme.
Article
Biotechnology & Applied Microbiology
Yiran Qu, Innocent Bekard, Ben Hunt, Jamie Black, Louis Fabri, Sally L. Gras, Sandra. E. Kentish
Summary: This study compares the performance of a nanofiber device and a resin column for antibody capture. The nanofiber device has a larger housing volume and lower binding capacity, but comparable eluate purity to the resin column. It shows high stability, can be used for multiple cycles, and maintains consistent eluate quality when scaled up. The use of a single nanofiber device can significantly reduce costs compared to a resin column, especially when the number of batches is limited.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Brandon Moore, Christos Georgakis, Chris Antoniou, Sarwat Khattak
Summary: Fed-batch cell culture processes are commonly used in biomanufacturing due to their simplicity and applicability in cGMP environments. However, the challenge lies in the changing physiochemical conditions within the bioreactor as the cell density changes. Traditional response surface models (RSMs) are commonly used for optimization but are limited by their use of time-invariant factors. Dynamic RSM (DRSM) models can predict the time-dependent impact of process inputs, allowing for optimization of process operations that change over time.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Lin Li, Yunfan Bai, Chuhua Qi, Yile Du, Xiaoxiao Ma, Yutong Li, Pingping Wu, Shuangli Chen, Sijing Zhang
Summary: A succinic anhydride-modified apple pomace (SAMAP) was synthesized to address environmental issues caused by the accumulation of apple pomace and effectively treat heavy metal ions. SAMAP exhibited high adsorption capacity for Cu(II) and Pb(II), suggesting its potential application in wastewater treatment.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Peter Satzer
Summary: Water for injection (WFI) production in the biopharmaceutical industry consumes excessive amounts of water and energy. Recycling buffers can potentially save up to 90% of resources, but achieving the full theoretical potential is impossible when a risk-aware design is used. Universal risk-based assessment is important for regulatory authorities to consider the implementation of such a strategy.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Gaoya Sun, Lingkai Jin, Guangxue Wang, Xiaoge Wang, Jin Huang
Summary: In this study, heterologous expression and homologous overexpression of ABC transporter proteins AatA and MdlB were found to improve butyric acid production in C. tyrobutyricum. The overexpression of these proteins upregulated the expression levels of key enzymes in the acetate synthesis pathway and promoted the synthesis and secretion of acetic acid. Additionally, the increase in ATPase activity facilitated sugar utilization, induced extracellular secretion of acetate, and shortened fermentation periods.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Taiki Okamura, Rina Aritomi, Takuya Matsumoto, Ryosuke Yamada, Hidehiko Hirakawa, Hiroyasu Ogino
Summary: In this study, proline was introduced to improve the stability of putidaredoxin reductase (PdR) in the Pseudomonas putida cytochrome P450 system. It was found that PdR_T221P had a longer half-life at high temperatures compared to wild-type PdR, but a shorter half-life in the presence of methanol. Molecular dynamics simulations supported these findings.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Yuying Hu, Xiaofan Wang, Shihao Zhang, Zimu Liu, Tengfang Hu, Xin Wang, Xiaoming Peng, Hongling Dai, Jing Wu, Fengping Hu
Summary: This study investigates the effect of iron-carbon micro-electrolysis (ICME) materials on high-solid anaerobic digestion (HSAD). The results show that ICME materials promote methane production in HSAD by increasing the attachment area of microorganisms and facilitating symbiotic metabolism of certain bacterial species. This study provides new insights into microbial mechanisms and enhances our understanding of ICME material enhancement in HSAD.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Jibao Liu, Yufeng Xu, Yuansong Wei
Summary: This study investigated the role of sludge rheology in anaerobic digestion (AD) and found that rheological properties increased with the increase of solid content, resulting in a negative effect on methane production. An extended ADM1 model revealed that enhanced sludge rheological properties increased mass diffusion resistance and reduced uptake rate of acetate.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Xiaoyan Liu, Zongze Chen, Dewen Kong, Xinying Zhang, Chuanhua Wang, Yongqi Wang
Summary: This study explored the role of intracellular and extracellular enzymes of Acinetobacter baumannii and Talaromyces sp. in the degradation of crude oil. The extracellular enzymes of Talaromyces sp. were more effective in degrading n-alkanes, while those of Acinetobacter baumannii had a better effect on aromatic hydrocarbons. The degradation enzyme systems of both bacteria and fungi complemented each other, improving the overall degradation ability.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Jing Dong, Lingli Xu, Yuxiang Liu, Li Ren, Ke Yuan
Summary: The utilization of biochar-immobilized microorganisms is an effective method for eliminating phenol from water. The high susceptibility of bacteria to environmental factors is a challenge for practical implementation. In this study, biochar was used to reduce microbial susceptibility and enhance phenol removal. The addition of biochar altered the dominant species of phenol-degrading bacteria and response surface analysis indicated the significant influence of biochar pyrolysis temperature and experimental temperature on phenol removal rate.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Zibin Pan, Mengying Liu, Zuliang Chen
Summary: This study successfully removed metalloids and heavy metals from acid mine drainage (AMD) using bio-synthesized Fe/Cu nanoparticles (Fe/Cu NPs). The Fe/Cu NPs showed high removal capacities and the presence of organic substances contributed to their stability.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Guangbing Liu, Han Zhang, Jincan Huang, Lu Zhang, Teng Zhang, Xuemin Yu, Weijing Liu, Chunkai Huang
Summary: This study investigated the effect of Fenton pre-treatment on the treatment efficiency of printing and dyeing wastewater (PDW) using two anaerobic/aerobic-membrane bioreactors (A/O-MBRs). The results showed that Fenton pre-treatment significantly improved the removal efficiency of COD and AOX in PDW, and reduced membrane fouling. The Shannon indices and metagenomics analysis indicated that the microbial diversity in anaerobic flocs was higher than that in aerobic flocs, and EC3.1.1.45 and pcaI were identified as key functional genes.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Lidia Borgosz, Duygu Dikicioglu
Summary: The Industrial Internet of Things (IIoT) is a system that connects devices and provides real-time insight into industrial processes. However, the complexity and regulatory requirements of the biomanufacturing sector make it challenging to implement IIoT. There is a need for universal solutions to overcome this challenge and advance the field of biomanufacturing.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Yajie Li, Weikang Kong, Yuyao Zhang, Huarui Zhou, Hongbo Liu, Salma Tabassum
Summary: In this study, the iron-carbon multi-micro electric field coupling anaerobic co-digestion technique was used to treat coal gasification wastewater (CGW). The experimental results showed that under optimal operating conditions, this technique can significantly reduce the toxicity of the wastewater and achieve high removal efficiencies. Additionally, the analysis of microbial communities revealed that the coupling system promotes direct interspecies electron transfer.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
Article
Biotechnology & Applied Microbiology
Aparecido Nivaldo Modenes, Debora Gozzi Fernandes, Daniela Estelita Goes Trigueros, Matheus Guilherme Amador, Fernando Rodolfo Espinoza-Quinones, Taysa de Souza Braniz, Adilson Ricken Schuelter, Glacy Jaqueline da Silva, Lucimar Pereira Bonett
Summary: This study aimed to systematically remove organic pollutants from raw dairy wastewater with high concentrations of COD, TOC, and TN using Poterioochromonas malhamensis algae strains. The results showed that the biomass yield rate using FP-PBRs was 10% higher than tubular PBRs, and the organic pollution in wastewater was significantly reduced with a decrease of about 98% in COD, 95% in TN, and 92% in TOC.
BIOCHEMICAL ENGINEERING JOURNAL
(2024)
