Article
Biotechnology & Applied Microbiology
Sean F. Sullivan, Anuj Shetty, Tharun Bharadwaj, Naveen Krishna, Vikas D. Trivedi, Venkatesh Endalur Gopinarayanan, Todd C. Chappell, Daniel M. Sellers, R. Pravin Kumar, Nikhil U. Nair
Summary: Engineering the utilization of non-native substrates in industrial microbes provides a way to use renewable sources for bioprocesses. Previous research has shown that activating the GAL regulon in Saccharomyces cerevisiae during growth on the non-native substrate xylose enhances gene expression and growth compared to constitutive overexpression of the same pathway. However, this method was not easily adaptable to other non-native substrates. In this study, a variant of Gal3p called Gal3pMC was constructed, which can activate the GAL regulon with diverse substrates and mimic the dynamics of the native system. Testing this regulon approach with non-native lignocellulosic sugars resulted in higher growth rates and final cell densities compared to constitutive overexpression. The co-utilization of multiple non-native substrates suggests that this regulon approach could be universally beneficial for engineering synthetic heterotrophy.
METABOLIC ENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Irene Otero-Muras, Pablo Carbonell
Summary: Metabolic engineering involves optimizing processes from single-cell to fermentation to increase production of valuable chemicals. A systems approach has accelerated scaling from rapid prototyping to industrial production, with automated DNA assembly reducing time from conception to production. The success of metabolic engineering often relies on retrobiosynthetic protocols and dynamic regulation strategies assembled as genetic circuits in host strains.
METABOLIC ENGINEERING
(2021)
Review
Biochemistry & Molecular Biology
Dongsoo Yang, Hyunmin Eun, Cindy Pricilia Surya Prabowo
Summary: Polyketides, a type of natural products, have versatile applications such as pharmaceuticals, nutraceuticals, and cosmetics. Aromatic polyketides, including type II and III polyketides, are important chemicals for human health, such as antibiotics and anticancer agents. However, they are mainly produced from challenging sources like soil bacteria or plants. To address this issue, metabolic engineering and synthetic biology have been used to efficiently engineer model microorganisms for enhanced production of aromatic polyketides. This review discusses the recent advancements and future prospects of metabolic engineering and synthetic biology strategies in the production of type II and type III polyketides in model microorganisms.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Editorial Material
Biochemical Research Methods
Michael B. Sheets, Joshua T. Atkinson, Mark P. Styczynski, Emily R. Aurand
Summary: As engineering biology continues to have increasing impacts, it is crucial to introduce the field early on in an accessible manner. However, teaching engineering biology faces challenges such as limited representation in widely used scientific textbooks or curricula, as well as its interdisciplinary nature. To address this, we have developed an adaptable curriculum module that provides a versatile slide deck designed by engineering biology experts to cover the fundamental principles and applications of the field. Free and accessible through a public website, this module can be used independently or integrated into existing curricula, aiming to enhance the ease of teaching current engineering biology topics and promote public engagement with the field.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Ana P. Teixeira, Pascal Stucheli, Simon Auslander, David Auslander, Pascal Schonenberger, Samuel Hurlemann, Martin Fussenegger
Summary: The researchers have developed an antibiotic-free selection system for generating stable transgenic cell lines, which utilizes cellobiose as the primary energy source. By combining this system with a PiggyBac transposase-based integration strategy, they were able to efficiently generate stable transgenic cell lines that expressed high levels of cargo proteins. This strategy was successfully validated by integrating sequences encoding two biopharmaceutical proteins, erythropoietin and the monoclonal antibody rituximab, and confirmed their efficient production in both cellobiose- and glucose-containing medium.
METABOLIC ENGINEERING
(2022)
Article
Biotechnology & Applied Microbiology
Shengyu Li, Pengzhi Zhang, Weiqing Chen, Lingqun Ye, Kristopher W. Brannan, Nhat-Tu Le, Jun-ichi Abe, John P. Cooke, Guangyu Wang
Summary: RNA velocity offers a method to infer cell state transitions from single-cell RNA sequencing data. cellDancer is a scalable deep neural network that locally infers velocity for each cell and provides single-cell resolution inference of velocity kinetics. It shows robust performance in multiple kinetic regimes, high dropout ratio datasets, and sparse datasets. Additionally, cellDancer provides cell-specific predictions of transcription, splicing, and degradation rates, which are potential indicators of cell fate.
NATURE BIOTECHNOLOGY
(2023)
Review
Biotechnology & Applied Microbiology
Govinda R. Navale, Mahesh S. Dharne, Sandip S. Shinde
Summary: Isoprenoids, also known as terpenoids, are diverse organic compounds found abundantly in nature, with various applications in pharmaceutical, nutraceutical, and chemical industries. Recent advancements in metabolic engineering have enabled the production of isoprenoids in heterologous host systems like Escherichia coli and Saccharomyces cerevisiae. Challenges and strategies for scale-up and engineering of isoprenoids in heterologous host systems have been discussed in detail.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Review
Plant Sciences
Revuru Bharadwaj, Sarma R. Kumar, Ashutosh Sharma, Ramalingam Sathishkumar
Summary: Gene clusters responsible for specialized metabolites in plants offer a potential untapped source for highly useful biomolecules. These clusters evolve through gene duplications and neofunctionalization, tightly regulated at nucleosome level. Further research is needed to explore the significance and regulatory mechanisms of these gene clusters in plant biosynthesis.
FRONTIERS IN PLANT SCIENCE
(2021)
Review
Plant Sciences
Revuru Bharadwaj, Sarma R. Kumar, Ashutosh Sharma, Ramalingam Sathishkumar
Summary: Plants contain gene clusters encoding enzymes for the biosynthesis of specialized metabolites, which have evolved through gene duplications and neofunctionalization and are tightly regulated at nucleosome level. The prevalence of these gene clusters presents an attractive possibility of untapped source of highly useful biomolecules in plants.
FRONTIERS IN PLANT SCIENCE
(2021)
Article
Pharmacology & Pharmacy
Rita P. Lopes, Raquel A. Ferro, Margarida Milhazes, Margarida Figueira, Maria Joao Caldeira, Alexandra M. M. Antunes, Helena Gaspar
Summary: Synthetic cathinones, including both reported and unreported substances, are widely used for recreational purposes, posing a serious public health issue. It is challenging for forensic authorities to keep up with the constantly changing NPS market. The lack of pharmacokinetic data further complicates forensic and clinical verifications.
FRONTIERS IN PHARMACOLOGY
(2023)
Review
Plant Sciences
Vimalraj Mani, Soyoung Park, Jin A. Kim, Soo In Lee, Kijong Lee
Summary: Terpenoids, with vast structural diversity, are high-value specialized metabolites widely used in cosmetics, foods, and pharmaceuticals. Recent advancements in heterologous production platforms and metabolic engineering technologies have overcome challenges related to the bioavailability of terpenoids in natural sources. Studies have shown that subcellular localization of precursor pool and introduced enzymes are crucial for increasing targeted terpenoid production in plants.
Review
Biotechnology & Applied Microbiology
Bhagyashree Bachhav, Jacopo de Rossi, Carlos D. Llanos, Laura Segatori
Summary: The production of high-quality recombinant proteins is crucial for the continuous supply of biopharmaceuticals, such as therapeutic antibodies. Engineering mammalian cell factories faces limitations due to proteotoxic stress caused by abnormal accumulation of protein folding intermediates, leading to apoptosis. This review discusses advances in cell engineering, focusing on controlling the expression of recombinant proteins at various levels and highlights the opportunities to use synthetic biology tools for designing programmable cell factories for improved biomanufacturing of therapeutic proteins.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Review
Biotechnology & Applied Microbiology
Javad Aminian-Dehkordi, Shadi Rahimi, Mehdi Golzar-Ahmadi, Amritpal Singh, Javiera Lopez, Rodrigo Ledesma-Amaro, Ivan Mijakovic
Summary: Synthetic biology offers new solutions for environmental protection by developing remediation systems using genetically engineered microbes and plants, and utilizing computational methods for design and application to detect and respond to specific pollutants.
BIOTECHNOLOGY ADVANCES
(2023)
Review
Biology
Almando Geraldi, Fatiha Khairunnisa, Nadya Farah, Le Minh Bui, Ziaur Rahman
Summary: Biotechnology uses biological synthesis to create high-value products, with metabolic engineering adjusting cellular pathways to enhance production. Biological scaffolds anchor enzymes and substrates to improve production by fixing them in the right location and orientation, serving as a strategy to address challenges in achieving efficient metabolic activities.
Review
Biotechnology & Applied Microbiology
Andrew Currin, Steven Parker, Christopher J. Robinson, Eriko Takano, Nigel S. Scrutton, Rainer Breitling
Summary: The ability to engineer biological systems is crucial in biotechnology and synthetic biology. Generating genetic diversity is essential to explore variations in phenotype, and recent advancements in DNA synthesis technology have enhanced our ability to create variant libraries. This review focuses on approaches to create diversity from enzyme level to entire pathways in vitro, emphasizing the creation of combinatorial libraries for significant improvements in function.
BIOTECHNOLOGY ADVANCES
(2021)
Review
Biotechnology & Applied Microbiology
Christopher J. Hartline, Alexander C. Schmitz, Yichao Han, Fuzhong Zhang
Summary: Metabolic engineering enables the production of valuable chemicals using microbial organisms, with dynamic metabolic engineering allowing cells to autonomously adjust flux in response to their metabolic state. The review discusses theoretical works, molecular mechanisms, and applications of dynamic control systems in improving metrics in metabolic engineering.
METABOLIC ENGINEERING
(2021)
Article
Biotechnology & Applied Microbiology
Alexander Schmitz, Fuzhong Zhang
Summary: Our study demonstrates that sequences with higher tRNA Adaptation Index (TAI) scores and Codon Adaptation Index (CAI) scores exhibit higher variation in green fluorescent protein (GFP) expression. This variation is not observed in sequences with high Normalized Translation Efficiency Index (nTE) scores or based on mRNA secondary structure folding energy. We found that noise in GFP expression scales with mean protein abundance for low-abundant proteins but remains constant for high-abundant proteins, suggesting that noise for high-abundance proteins is not primarily due to translation elongation. This indicates that codon optimization can be performed without worrying about gene expression noise in biotechnology applications.
Article
Chemistry, Multidisciplinary
Jingyao Li, Yaguang Zhu, Han Yu, Bin Dai, Young-Shin Jun, Fuzhong Zhang
Summary: Researchers have developed a new strategy for synthesizing amyloid proteins by fusing amyloid peptides with flexible linkers from spider silk proteins, resulting in fibers with strong mechanical properties. These fibers can be utilized for a variety of mechanically demanding applications, surpassing most recombinant protein fibers and even some natural spider silk fibers in terms of ultimate tensile strength and toughness.
Article
Biochemical Research Methods
Babita K. Verma, Ahmad A. Mannan, Fuzhong Zhang, Diego A. Oyarzun
Summary: Recent advances in synthetic biology have enabled the construction of dynamic control circuits for metabolic engineering, which can self-regulate gene expression in response to bioreactor perturbations. The central components in these circuits are metabolite biosensors, but their construction remains a bottleneck in strain design.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Microbiology
Christopher J. Hartline, Ruixue Zhang, Fuzhong Zhang
Summary: This study found that shifts in carbon sources and positive feedback in nutrient transport are not enough to trigger persistence in the majority of bacteria, but instead only lead to temporary tolerance. Additionally, the duration of this temporary tolerance is determined by the metabolic state prior to the shift, and supplying glyoxylate can facilitate antibiotic killing of bacteria.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biochemical Research Methods
Christopher J. Hartline, Fuzhong Zhang
Summary: Metabolite biosensors based on metabolite-responsive transcription factors are important components in synthetic biology for sensing and controlling cellular metabolism. This study investigates the impact of growth rate on biosensor performance and reveals that different biosensors exhibit different growth rate dependencies in their dynamic range. The study also develops a kinetic model to explore the effects of tuning biosensor parameters on dynamic range and its growth rate dependence.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Wenqin Bai, Winston E. Anthony, Christopher J. Hartline, Shaojie Wang, Bin Wang, Jie Ning, Fong-Fu Hsu, Gautam Dantas, Fuzhong Zhang
Summary: Bacterial fatty acids are essential components of cellular membranes and important sources of renewable chemicals. Modifying their structure can expand their applications and affect the growth and metabolic state of the producing cells. This study successfully engineered Escherichia coli to have different fatty acid profiles, and found that these profiles had varying effects on transcriptome, growth, and stress response.
METABOLIC ENGINEERING
(2022)
Review
Biochemistry & Molecular Biology
Juya Jeon, Shri Venkatesh Subramani, Kok Zhi Lee, Bojing Jiang, Fuzhong Zhang
Summary: High molecular weight (MW), highly repetitive protein polymers have the potential to replace petroleum-derived materials due to their renewable, biodegradable nature and exceptional mechanical properties. However, their synthesis in large quantities for real-world applications is challenging due to their high MW and repetitive sequence features. To address this challenge, various methods, including construction of repetitive genes, expression of repetitive proteins from circular mRNAs, and synthesis of repetitive proteins through ligation and protein polymerization, have been developed. This review discusses the advantages, limitations, and future directions for the scalable production of highly repetitive protein polymers for diverse applications.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Multidisciplinary Sciences
Jingyao Li, Bojing Jiang, Xinyuan Chang, Han Yu, Yichao Han, Fuzhong Zhang
Summary: The authors have developed a method to enhance the strength of low molecular-weight protein materials by fusing intrinsically-disordered mussel foot protein fragments. This approach can be applied to a wide range of protein-based materials and has the potential to achieve high yields.
NATURE COMMUNICATIONS
(2023)
Review
Chemistry, Analytical
Gloria J. Zhou, Fuzhong Zhang
Summary: TF-based biosensors are commonly used for metabolite detection and cellular pathway regulation. However, their direct application to new hosts or metabolic pathways often requires extensive tuning for optimal performance. This review highlights recent strategies for engineering TF-based biosensors to achieve desired performance and discusses additional design considerations. The article also examines applications of these sensors and suggests important areas for further research.
Review
Biotechnology & Applied Microbiology
Xinyue Mu, Fuzhong Zhang
Summary: This article provides a comprehensive overview of heterogeneity in microbial bioproduction, including its mechanisms and strategies for control. By understanding and controlling heterogeneity, economic competitiveness of biomanufacturing can be enhanced.
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Po-Cheng Lin, Fuzhong Zhang, Himadri B. Pakrasi
Summary: Terpenoids are a diverse group of natural products with commercial applications, and microbial production of terpenes in cyanobacteria is a promising approach. This study successfully engineered a cyanobacterium to produce limonene by identifying a beneficial mutation in a key gene, leading to a significant increase in production. Combinatorial metabolic engineering was used to optimize gene expression levels in the biosynthesis pathway, demonstrating the importance of fine-tuning gene expression for enhancing terpene production in cyanobacteria.
METABOLIC ENGINEERING COMMUNICATIONS
(2021)