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Article
| Open AccessWax worm saliva and the enzymes therein are the key to polyethylene degradation by Galleria mellonella
The crucial first step in the biodegradation of polyethylene plastic is oxidation of the polymer. This has traditionally required abiotic pre-treatment, but now Bertocchini and colleagues report two wax worm enzymes capable of catalyzing this oxidation and subsequent degradation at room temperature.
- A. Sanluis-Verdes
- , P. Colomer-Vidal
- & F. Bertocchini
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Article
| Open AccessA miniaturized bionic ocean-battery mimicking the structure of marine microbial ecosystems
Marine ecosystems are dominated by microbial communities. Inspired by the photoelectric conversion feature of the marine ecosystems, the authors design a four-species microbial community to mimic primary producer, primary degrader, and ultimate consumers, and show its ability to convert light into electricity for over one month.
- Huawei Zhu
- , Liru Xu
- & Yin Li
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Article
| Open AccessInducible expression of large gRNA arrays for multiplexed CRISPRai applications
CRISPR gene activation and inhibition has become a powerful synthetic tool for influencing the expression of native genes for foundational studies, cellular reprograming, and metabolic engineering. Here the authors demonstrate near leak-free, inducible expression of a polycistronic array containing up to 24 gRNAs from two orthogonal CRISPR/Cas systems.
- William M. Shaw
- , Lucie Studená
- & Rodrigo Ledesma-Amaro
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Article
| Open AccessMuconic acid production from glucose and xylose in Pseudomonas putida via evolution and metabolic engineering
Muconic acid is a platform chemical with wide industrial applicability. Here, the authors report efficient muconate production from glucose and xylose by engineered Pseudomonas putida strain using adaptive laboratory evolution, metabolic modeling, and rational strain engineering strategies.
- Chen Ling
- , George L. Peabody
- & Gregg T. Beckham
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Article
| Open AccessAn in vivo gene amplification system for high level expression in Saccharomyces cerevisiae
Gene dosage-based expression upregulation suffers from instability and random gene integration. Here, the authors report HapAmp, a method that uses haploinsufficiency as evolutionary force to drive in vivo gene amplification, and demonstrate its applications in protein and biochemical production in yeast.
- Bingyin Peng
- , Lygie Esquirol
- & Claudia E. Vickers
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Article
| Open AccessUnlocking the functional potential of polyploid yeasts
Domesticated industrial yeast strains are sterile, which hampers to breed strains with novel properties. Here, the authors employ the genetics paradigm return-to-growth to induce genome wide recombination in two sterile polyploid industrial yeasts and identify clones with superior biotechnological traits.
- Simone Mozzachiodi
- , Kristoffer Krogerus
- & Gianni Liti
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Article
| Open AccessEnhancing biofuels production by engineering the actin cytoskeleton in Saccharomyces cerevisiae
Product toxicity is one of the factors that hinder biofuel production. Here, the authors engineer the actin cytoskeleton to increase cell growth and production of n-butanol and medium-chain fatty acids in Saccharomyces cerevisiae.
- Hui Liu
- , Pei Zhou
- & Liming Liu
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Article
| Open AccessMiDAS 4: A global catalogue of full-length 16S rRNA gene sequences and taxonomy for studies of bacterial communities in wastewater treatment plants
Microbial communities are responsible for biological wastewater treatment. Here, Dueholm et al. generate more than 5 million high-quality, full-length 16S rRNA gene sequences from wastewater treatment plants across the world to construct a database with a comprehensive taxonomy, providing insights into diversity and function of these microbial communities.
- Morten Kam Dahl Dueholm
- , Marta Nierychlo
- & Per Halkjær Nielsen
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Article
| Open AccessExploring a blue-light-sensing transcription factor to double the peak productivity of oil in Nannochloropsis oceanica
Microalgae are promising feedstock for oil production. The authors report that a transcription factor NobZIP77 can regulate oil synthesis by sensing the blue light, and explore these findings to greatly enhance oil productivity via genetic and process engineering in Nannochloropsis oceanica.
- Peng Zhang
- , Yi Xin
- & Jian Xu
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Article
| Open AccessMetabolic engineering strategies to produce medium-chain oleochemicals via acyl-ACP:CoA transacylase activity
Microbial production of oleochemicals involves strategies of expressing thioesterase to narrow the substrate pool for the termination enzyme at the expense of one ATP. Here, the authors developed an alternative energy-efficient strategy to use of an acyl-ACP transacylase to produce medium chain oleochemicals in E. coli.
- Qiang Yan
- , William T. Cordell
- & Brian F. Pfleger
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Article
| Open AccessSynthetic glycans control gut microbiome structure and mitigate colitis in mice
Here, the authors characterize the gut microbiome fermentation properties and therapeutic potential of chemically diverse synthetic glycans (SGs), showing they promote specific shifts in taxonomic and metabolite profiles, and exhibit therapeutic benefits in mouse models of colonic inflammation, together implying SGs as a potential avenue to treat disease by modulating the composition and metabolites produced by the gut microbiome.
- Andrew C. Tolonen
- , Nicholas Beauchemin
- & Johan E. T. van Hylckama Vlieg
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Article
| Open AccessCRISPR-assisted rational flux-tuning and arrayed CRISPRi screening of an l-proline exporter for l-proline hyperproduction
Corynebacterium glutamicum is a major workhorse in industrial biomanufacturing of amino acids. Here, the authors employ CRISPR-assisted rational flux-tuning and CRISPRi screening of a L-proline exporter to covert a wild-type C. glutamicum to a hyperproducer of L-proline.
- Jiao Liu
- , Moshi Liu
- & Jibin Sun
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Article
| Open AccessMachine learning-informed and synthetic biology-enabled semi-continuous algal cultivation to unleash renewable fuel productivity
Growth limitation caused by mutual shading and the high harvest cost hamper algal biofuel production. Here, the authors overcome these two problems by designing a semi-continuous algal cultivation system and an aggregation-based sedimentation strategy to achieve high levels production of biomass and limonene.
- Bin Long
- , Bart Fischer
- & Joshua S. Yuan
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Article
| Open AccessMapping enzyme catalysis with metabolic biosensing
The testing of engineered enzymes represents a bottleneck. Here the authors report a screening method combining microfluidics and mass spectrometry, to map the catalysis of a mutated enzyme, characterise the range of products generated and recover the sequences of variants with desired activities.
- Linfeng Xu
- , Kai-Chun Chang
- & Adam R. Abate
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Article
| Open AccessDesigning the bioproduction of Martian rocket propellant via a biotechnology-enabled in situ resource utilization strategy
Returning from Mars to Earth requires propellant. The authors propose a biotechnology-enabled in situ resource utilization (bioISRU) process to produce a Mars specific rocket propellant, 2,3-butanediol, using cyanobacteria and engineered E. coli, with lower payload mass and energy usage compared to chemical ISRU strategies.
- Nicholas S. Kruyer
- , Matthew J. Realff
- & Pamela Peralta-Yahya
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Article
| Open AccessMicrobial production of megadalton titin yields fibers with advantageous mechanical properties
Here, the authors engineer microbial production of muscle titin fibers with highly desirable mechanical properties and provide structural analyses that explain the molecular mechanisms underlying high performance of this polymer with potential uses in biomedicine and textile industries, among others.
- Christopher H. Bowen
- , Cameron J. Sargent
- & Fuzhong Zhang
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Article
| Open AccessComplete and efficient conversion of plant cell wall hemicellulose into high-value bioproducts by engineered yeast
Cellulosic hydrolysates contain substantial amounts of acetate, which is toxic to fermenting microorganisms. Here, the authors engineer Baker’s yeast to co-consume xylose and acetate for triacetic acid lactone production from a hemicellulose hydrolysate of switchgrass.
- Liang Sun
- , Jae Won Lee
- & Yong-Su Jin
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Article
| Open AccessMetabolome and proteome analyses reveal transcriptional misregulation in glycolysis of engineered E. coli
Synthetic pathways represent a metabolic burden on host cells. Here the authors engineer Cra-binding sites to prevent misregulation in glycerol and carotenoid overproducing E. coli strains.
- Chun-Ying Wang
- , Martin Lempp
- & Hannes Link
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Article
| Open AccessRenewable fatty acid ester production in Clostridium
Esters can be used as fuels and specialty chemicals for food flavoring, cosmetic and pharmaceutical industries. Here, the authors systematically engineer clostridia, including discovery and deletion of prophages to increase strain stability, for the production of butyl acetate and butyl butyrate from corn stover at low cost.
- Jun Feng
- , Jie Zhang
- & Yi Wang
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Article
| Open AccessCreating enzymes and self-sufficient cells for biosynthesis of the non-natural cofactor nicotinamide cytosine dinucleotide
Nicotinamide cytosine dinucleotide is an NAD mimic that can be used to mediate redox reactions. Here the authors design an NCD synthetase for the intracellular production of NCD.
- Xueying Wang
- , Yanbin Feng
- & Zongbao K. Zhao
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Article
| Open AccessHyperproduction of 3-hydroxypropionate by Halomonas bluephagenesis
3-Hydroxypropionic acid (3HP) is an important platform chemical. Here, the authors engineer Halomonas bluephagenesis by deleting newly identified degradation pathway and balancing redox state to achieve high level production of 3HP and its copolymer under open and unsterile conditions.
- Xiao-Ran Jiang
- , Xu Yan
- & Guo-Qiang Chen
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Article
| Open AccessComplex yeast–bacteria interactions affect the yield of industrial ethanol fermentation
Industrial sugarcane ethanol fermentations are accomplished by a microbial community dominated by S. cerevisiae and co-occurring bacteria. Here, the authors investigate how microbial community composition contributes to community function and reveal the role of acetaldehyde in improving yeast growth rate and ethanol production.
- Felipe Senne de Oliveira Lino
- , Djordje Bajic
- & Morten Otto Alexander Sommer
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Article
| Open AccessEngineered yeast genomes accurately assembled from pure and mixed samples
The cost and complexity of whole genome sequencing limits its use in identifying and validating sequences used for genetic engineering and synthetic biology. Here the authors present Prymetime, an integrated workflow to sequence engineered strains and identify engineering in metagenomes.
- Joseph H. Collins
- , Kevin W. Keating
- & Eric M. Young
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Article
| Open AccessComplete biosynthesis of a sulfated chondroitin in Escherichia coli
Chondroitin sulfate (CS) is a type of sulfated glycosaminoglycan that is manufactured by extraction from animal tissues for the treatment of osteoarthritis and in drug delivery applications. Here, the authors report the development of single microbial cell factories capable of compete, one-step biosynthesis of animal-free CS production in E. coli.
- Abinaya Badri
- , Asher Williams
- & Mattheos A. G. Koffas
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Article
| Open AccessCas12a-assisted precise targeted cloning using in vivo Cre-lox recombination
Direct cloning of biosynthetic gene clusters is difficult due to their size and repetitive nature. Here, the authors present CAPTURE, which uses Cas12a and in vivo Cre-lox recombination for efficient cloning of gene clusters up to 113 kb, and demonstrate how this method can be used for large-scale discovery of novel natural products.
- Behnam Enghiad
- , Chunshuai Huang
- & Huimin Zhao
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Article
| Open AccessAuxin-mediated protein depletion for metabolic engineering in terpene-producing yeast
Loss-of-function experiments are used in metabolic engineering to understand and optimise metabolism. Here, the authors apply auxin inducible protein degradation to test different metabolic engineering strategies for improved terpenoid production in yeast.
- Zeyu Lu
- , Bingyin Peng
- & Claudia E. Vickers
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Article
| Open AccessMicrobial production of multiple short-chain primary amines via retrobiosynthesis
Short-chain primary amines (SCPAs) are industrially important compounds that are commonly produced under harsh synthetic conditions. Here, the authors report a combination of retrobiosynthesis and precursor selection step for design of biosynthetic pathways leading to production of SCPAs, using valine decarboxylase-expressing Escherichia coli strains.
- Dong In Kim
- , Tong Un Chae
- & Sang Yup Lee
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Article
| Open AccessSynthesis of high-titer alka(e)nes in Yarrowia lipolytica is enabled by a discovered mechanism
Alka(e)nes with chain lengths in C5-C23 range are ideal fuel components. Here, the authors report that high-titer production of alak(e)nes in pathway engineered Yarrowia lipolytica, which is enabled by the finding that acyl-CoA is another substrate of fatty acid photodecarboxylase (FAP).
- Jingbo Li
- , Yongshuo Ma
- & Gregory Stephanopoulos
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Article
| Open AccessPhotosynthetic hydrogen production by droplet-based microbial micro-reactors under aerobic conditions
The development of techniques capable of orchestrating the assembly of living cells into multicellular ensembles with synergistic and function is challenge. Here, the authors construct algal or algal/bacterial cells-based core shell-like structure based on aqueous two-phase system for synergic photosynthetic H2 production.
- Zhijun Xu
- , Shengliang Wang
- & Stephen Mann
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Article
| Open AccessProton-enabled activation of peptide materials for biological bimodal memory
The structural programmability and functionality of peptide materials can be leverage for various next-generation devices such as non-volatile memories. The authors report a proton-coupled mechanism in tyrosine-rich peptides for realizing multimodal memory devices.
- Min-Kyu Song
- , Seok Daniel Namgung
- & Jang-Yeon Kwon
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Article
| Open AccessSpace station biomining experiment demonstrates rare earth element extraction in microgravity and Mars gravity
Rare earth elements are used in electronics, but increase in demand could lead to low supply. Here the authors conduct experiments on the International Space Station and show microbes can extract rare elements from rocks at low gravity, a finding that could extend mining potential to other planets.
- Charles S. Cockell
- , Rosa Santomartino
- & René Demets
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Article
| Open AccessTitrating bacterial growth and chemical biosynthesis for efficient N-acetylglucosamine and N-acetylneuraminic acid bioproduction
An imbalance between cell growth and bioproduction of engineered microbes often reduces production efficiency. Here, the authors report genetic code expansion-based cell growth and biosynthesis balance engineering to achieve high levels production of N-acetylglucosamine in E. coli and N-acetylneuraminic acid in B. subtilis.
- Rongzhen Tian
- , Yanfeng Liu
- & Jian Chen
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Article
| Open AccessEngineering and application of a biosensor with focused ligand specificity
Transcriptional biosensors represent powerful tools for the screening of vast strain libraries, but the broad ligand specificity of some transcriptional regulators (TRs) can prohibit such applications. Here authors present the engineering of a LysG-based biosensor with a focused ligand specificity to isolate L-histidine-producing strains.
- Dennis Della Corte
- , Hugo L. van Beek
- & Jan Marienhagen
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Article
| Open AccessSynthetic microbial communities of heterotrophs and phototrophs facilitate sustainable growth
Successful application of microbial community for bioproduction relies on the selection of appropriate heterotroph and phototroph partners. Here, the authors construct community metabolic models to guide strain selection and experimentally validate metabolic exchanges that sustain the heterotrophs in minimal media.
- Cristal Zuñiga
- , Tingting Li
- & Karsten Zengler
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Article
| Open AccessRational flux-tuning of Halomonas bluephagenesis for co-production of bioplastic PHB and ectoine
Halomonas bluephagenesis is a halophilic platform bacterium for next generation industrial biotechnology. Here, the authors employ a stimulus response-based flux-tuning method for coproduction of bioplastic PHB and ectoine under open unsterile and continuous growth conditions.
- Hong Ma
- , Yiqing Zhao
- & Guo-Qiang Chen
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Article
| Open AccessEliminating the capsule-like layer to promote glucose uptake for hyaluronan production by engineered Corynebacterium glutamicum
Bioproduction of hyaluronan needs increases in yield and greater diversity of the molecular weights. Here, the author increases hyaluronan production and diversifies the molecular weights through engineering the hyaluronan biosynthesis pathway and disruption of Corynebacterium glutamicum encapsulation caused by secreted hyaluronan.
- Yang Wang
- , Litao Hu
- & Zhen Kang
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Article
| Open AccessLight-powered Escherichia coli cell division for chemical production
Manipulation of genes controlling microbial shapes can affect bio-production. Here, the authors employ an optogenetic method to realize dynamic morphological engineering of E. coli replication and division and show the increased production of acetoin and poly(lactate-co-3-hydroxybutyrate).
- Qiang Ding
- , Danlei Ma
- & Xiulai Chen
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Article
| Open AccessStress-induced expression is enriched for evolutionarily young genes in diverse budding yeasts
Fermentation parameters of industrial processes are often not the ideal growth conditions for industrial microbes. Here, the authors reveal that young genes are more responsive to environmental stress than ancient genes using a new gene age assignment method and provide targeted genes for metabolic engineering.
- Tyler W. Doughty
- , Iván Domenzain
- & John P. Morrissey
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Article
| Open AccessEnhanced succinic acid production by Mannheimia employing optimal malate dehydrogenase
Malate dehydrogenase (MDH) is one of the key enzymes for succinic acid (SA) bioproduction. Here, the authors report biochemical and structural analyses of various MDHs to reveal amino acids influencing the specific activity and susceptibility to substrate inhibition, and achieve industrial-level SA production.
- Jung Ho Ahn
- , Hogyun Seo
- & Sang Yup Lee
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Article
| Open AccessDevelopment of a thermophilic coculture for corn fiber conversion to ethanol
Corn fiber is a difficult feedstock to utilize due to its recalcitrant hemicellulose. Here, the authors characterize the recalcitrant structures, isolate a new bacterium to consume the hemicellulose, identify its enzymes, and show the benefit with increased conversion of corn fiber to ethanol.
- Dhananjay Beri
- , William S. York
- & Christopher D. Herring
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Article
| Open AccessSynthetic biology based construction of biological activity-related library of fungal decalin-containing diterpenoid pyrones
Combining genome mining and heterologous expression in a genetically tractable host can lead to bioactive natural products discovery and production. Here, the authors employ this strategy for new decalin-containing diterpenoid pyrenes production by expressing native, extended, and shunt pathways in Aspergillus oryzae.
- Kento Tsukada
- , Shono Shinki
- & Teigo Asai
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Article
| Open AccessPooled CRISPRi screening of the cyanobacterium Synechocystis sp PCC 6803 for enhanced industrial phenotypes
Developing cyanobacteria as CO2-neutral cell factories relies on the knowledge of the regulation mechanisms for growth and metabolism. Here, the authors develop an inducible CRISPRi gene repression library in Synechocystis sp. PCC 6803 and screens genes potentially affecting growth and L-lactate tolerance and production.
- Lun Yao
- , Kiyan Shabestary
- & Elton P. Hudson
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Article
| Open AccessA severe leakage of intermediates to shunt products in acarbose biosynthesis
Biosynthetic mechanism for the type 2 diabetes treatment drug acarbose is not fully revealed. Here, the authors show that shunt pathways and inefficient amino-deoxyhexose biosynthesis lead to 1-epi-valienol and valienol accumulation, and minimizing the flux to these shunt products can increase acarbose titer in Actinoplanes species.
- Qinqin Zhao
- , Yuchang Luo
- & Zixin Deng
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Article
| Open AccessA comparative genomics study of 23 Aspergillus species from section Flavi
Aspergillus fungi classified within the section Flavi include harmful and beneficial species. Here, Kjærbølling et al. analyse the genomes of 23 Flavi species, showing high genetic diversity and potential for synthesis of over 13,700 CAZymes and 1600 secondary metabolites.
- Inge Kjærbølling
- , Tammi Vesth
- & Mikael R. Andersen
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Article
| Open AccessTowards a fully automated algorithm driven platform for biosystems design
Existing efforts have been focused on one of the elements in the automation of the design, build, test, and learn (DBTL) cycle for biosystems design. Here, the authors integrate a robotic system with machine learning algorithms to fully automate the DBTL cycle and apply it in optimizing the lycopene biosynthetic pathway.
- Mohammad HamediRad
- , Ran Chao
- & Huimin Zhao
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Article
| Open AccessDesigning yeast as plant-like hyperaccumulators for heavy metals
Existing heavy metal bioremediation systems are mainly based on plants, which require long growing time in specific conditions. Here, the authors mimic the characteristics of plant hyperaccumulators to engineer more tractable baker’s yeast and achieve 10–100-fold higher accumulation of chromium, arsenic, or cadmium.
- George L. Sun
- , Erin. E. Reynolds
- & Angela M. Belcher
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Article
| Open AccessRewiring carbon metabolism in yeast for high level production of aromatic chemicals
Microbial production of aromatic amino acid (AAA)-derived chemicals remains an outstanding metabolic engineering challenge. Here, the authors engineer baker’s yeast for high levels p-coumaric acid production by rewiring the central carbon metabolism and channeling more flux to the AAA biosynthetic pathway.
- Quanli Liu
- , Tao Yu
- & Yun Chen
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Article
| Open AccessGalactose to tagatose isomerization at moderate temperatures with high conversion and productivity
Production of tagatose, a sugar substitute, by isomerization of galactose suffers from unfavorable enzymatic kinetics, low enzyme stability, and low equilibrium constant. Here, the authors simultaneously overcome these limitations by encapsulating l-arabinose isomerase in permeabilized Lactobacillus plantarum.
- Josef R. Bober
- & Nikhil U. Nair
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Article
| Open AccessDevelopment of a longevous two-species biophotovoltaics with constrained electron flow
Power densities of existing microbial biophotovoltaics (BPV) are low and unendurable. Here, the authors develop a BPV based on d-lactate mediated microbial consortium, which can generate an average power density of 135 mW·m−2 for over 40 days in a spatial-temporal separation setup with medium replenishment.
- Huawei Zhu
- , Hengkai Meng
- & Yin Li