Search

This study characterizes a cryptic (pro)phage-related sequence within the Caldibacillus debilis GB1 genome, designated CBP1.CBP1 is a Siphoviridae-like genome highly related to GBVS1 from Geobacillus sp. 6k51. The CBP1genome is a 37,315 bp region containing 69 putative ORFs with a GC content of 42% flanked on both sides by host DNA integrated into the main bacterial chromosome (contig 16). Bioinformatic analyses identified cassettes of genes within the CBP1 genome that were similar in function, yet distinct in sequence, from genes previously identified in GBVS1. All of CBP1 genes had less than 60% amino acid sequence identity with GBVS1by tBLASTx, with the exception of the TMP repeat gene. CBP1 possessed all the necessary genes to undergo a temperate/lytic phage life cycle, including excision, replication, structural genes, DNA packaging, and cell lyses. Proteomic analysis of CBP1 revealed the expression of 5 proteins. One of the expressed proteins was a transcriptional regulator protein homologous to the bacteriophage λ repressor protein (cI) expressed in high amounts from the CBP1 region, consistent with a lysogenic phage in a repressed state. The CBP1 protein expression profile during host growth provides unique insight into thermophilic Siphoviridae-like phages in the repressed state within their host cells. [ABSTRACT FROM AUTHOR]

Cellular membranes, predominantly described as a dynamic bilayer, are composed of different lipids, transmembrane proteins, and carbohydrates. Most research on biological membranes focuses on the identification, characterization, and mechanistic aspects of their different components. These studies provide a fundamental understanding of membrane structure, function, and dynamics, establishing a basis for the development of membrane engineering strategies. To date, approaches in this field concentrate on membrane adaptation to harsh conditions during industrial fermentation, which can be caused by temperature, osmotic, or organic solvent stress. With advances in the field of metabolic engineering and synthetic biology, recent breakthroughs include proof of concept microbial production of essential medicines, such as cannabinoids and vinblastine. However, long pathways, low yields, and host adaptation continue to pose challenges to the efficient scale up production of many important compounds. The lipid bilayer is profoundly linked to the activity of heterologous membrane-bound enzymes and transport of metabolites. Therefore, strategies for improving enzyme performance, facilitating pathway reconstruction, and enabling storage of products to increase the yields directly involve cellular membranes. At the forefront of membrane engineering research are re-emerging approaches in lipid research and synthetic biology that manipulate membrane size and composition and target lipid profiles across species. This review summarizes engineering strategies applied to cellular membranes and discusses the challenges and future perspectives, particularly with regards to their applications in host engineering and bioproduction. [Display omitted] • Membrane lipid diversity is a promising engineering target in biotechnology. • Membrane engineering enhances cell viability in response to bioprocessing stress. • Modulation of lipid composition improves performance of microbial cell factories. • Synthetic Biology strategies have emerged for membrane engineering. [ABSTRACT FROM AUTHOR]

• Mcl-PHA biosynthesis from different central metabolic routes was tested at low DO. • De novo mcl-PHA synthesis was not induced by oxygen limitation in P. putida LS46. • The content of monomers from β-oxidation intermediates was enriched at low DO. • The oxygen transfer rate could be used to control the content of certain monomers. Biosynthesis of medium chain length polyhydroxyalkanoates (mcl-PHAs) requires oxygen, but little is known of how mcl-PHA production is impacted in low dissolved oxygen (DO) environments, particularly when synthesized from monomers obtained via a de novo pathway. Growth and mcl-PHA synthesis characteristics under microaerophilic bioreactor conditions were observed in Pseudomonas putida LS46 when grown on carbon substrates requiring a de novo pathway, to varying extent, for mcl-PHA synthesis. Negligible PHA accumulated from glucose, acetic acid, or valeric acid under oxygen limitation (DO ≤ 0.15 mg L−1), while final mcl-PHA content reached 19.5% of cell dry mass (CDM) from hexanoic acid. The C6 content of the polymer obtained under low DO was 10% higher than in a DO excess (NH 4 -limited) control experiment, with less of the C10 monomers indicative of de novo synthesis. In all cases, the carbon-to-PHA yield was poor (< 0.1 C-mol C-mol−1). We conclude that, as opposed to utilizing β-oxidation intermediates, de novo biosynthesis of mcl-PHA using P. putida LS46 is not favorable in microaerophilic environments. In addition to scale-up implications, this work also suggests that lowering the bioreactor oxygen transfer rate can decrease the contribution of monomers incorporated using de novo synthesis for carbon feedstocks with mixed composition. [ABSTRACT FROM AUTHOR]

Aromatic alcohols (including β-phenylethanol, tyrosol and tryptophol) could contribute positively to the aroma flavor of huangjiu at an appropriate amount. In this study, it was found that the aromatic alcohols in huangjiu were predominantly synthesized by Saccharomyces cerevisiae HJ01. The Ehrlich pathway and shikimate pathway, including 31 genes, are responsible for aromatic alcohol biosynthesis in S. cerevisiae. During the first 120 h of huangjiu fermentation, the genes GAP1 HJ01 (permease), BAT2 HJ01 (transaminase), PDC1 HJ01 and PDC5 HJ01 (pyruvate decarboxylase), and ADH1 HJ01 (alcohol dehydrogenase) formed integrated Ehrlich pathway are responsible for aromatic alcohol biosynthesis. In S. cerevisiae HJ01, the mutations in the promoter region of genes GAP1 HJ01, BAT2 HJ01, PDC1 HJ01, and PDC5 HJ01 involved in the Ehrlich pathway increased the promoter strength and transcriptional levels of related genes. The addition of 20 U/g acid protease could substantially increase the expression levels of genes involved in the Ehrlich pathway, and therefore there were 34.7% increasing for the concentration of total aromatic alcohols. • Aromatic alcohols in huangjiu are synthesized by Saccharomyces cerevisiae. • Aromatic alcohols in huangjiu are mainly synthesized through the Ehrlich pathway. • Key genes and promoter mutations in Ehrlich pathway were identified. • Acid protease enhanced expression level of key genes. [ABSTRACT FROM AUTHOR]