Abstract:
BACKGROUND:Bacillus subtilis has been widely used as a host for heterologous protein expression in food industry. B. subtilis ATCC6051 is an alternative expression host for the production of industrial enzymes, and exhibits favorable growth properties compared to B. subtilis 168. Extracellular expression of pullulanase from recombinant B. subtilis is still limited due to the issues on promoters of B. subtilis expression system. This study was undertaken to develop a new, high-level expression system in B. subtilis ATCC6051. RESULTS:To further optimize B. subtilis ATCC6051 as a expression host, eight extracellular proteases (aprE, nprE, nprB, epr, mpr, bpr, vpr and wprA), the sigma factor F (spoIIAC) and a surfactin (srfAC) were deleted, yielding the mutant B. subtilis ATCC6051∆10. ATCC6051∆10 showed rapid growth and produced much more extracellular protein compared to the widetype strain ATCC6051, due to the inactivation of multiple proteases. Using this mutant as the host, eleven plasmids equipped with single promoters were constructed for recombinant expression of pullulanase (PUL) from Bacillus naganoensis. The plasmid containing the PspovG promoter produced the highest extracellular PUL activity, which achieved 412.9 U/mL. Subsequently, sixteen dual-promoter plasmids were constructed and evaluated using this same method. The plasmid containing the dual promoter PamyL-PspovG produced the maximum extracellular PUL activity (625.5 U/mL) and showed the highest expression level (the dry cell weight of 18.7 g/L). CONCLUSIONS:Taken together, we constructed an effective B. subtilis expression system by deleting multiple proteases and screening strong promoters. The dual-promoter PamyL-PspovG system was found to support superior expression of extracellular proteins in B. subtilis ATCC6051.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Liu X,Wang H,Wang B,Pan Ldoi
10.1186/s12934-018-1011-ysubject
Has Abstractpub_date
2018-10-22 00:00:00pages
163issue
1issn
1475-2859pii
10.1186/s12934-018-1011-yjournal_volume
17pub_type
杂志文章abstract:BACKGROUND:Extracts and products (roots and/or aerial parts) from Echinacea ssp. represent a profitable market sector for herbal medicines thanks to different functional features. Alkamides and polyacetylenes, phenols like caffeic acid and its derivatives, polysaccharides and glycoproteins are the main bioactive compou...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-44
更新日期:2013-05-04 00:00:00
abstract:BACKGROUND:In Saccharomyces cerevisiae, alpha-glucosidase (maltase) is a key enzyme in maltose metabolism. In addition, the overexpression of the alpha-glucosidase-encoding gene MAL62 has been shown to increase the freezing tolerance of yeast in lean dough. However, its cryoprotection mechanism is still not clear. RES...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01454-6
更新日期:2020-10-19 00:00:00
abstract::BACKGROUND: A rapid microtiter plate based sandwich hybridization assay was developed for detection and quantification of single RNA species using magnetic beads. Following solution hybridization target RNA molecules were collected by biotin-streptavidin affinity binding and detected by fluorescence signal generated b...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-2-4
更新日期:2003-04-28 00:00:00
abstract:BACKGROUND:Probiotic bacteria are increasingly used as immunomodulatory agents. Yet detailed molecular knowledge on the immunomodulatory molecules of these bacteria is lagging behind. Lipoteichoic acid (LTA) is considered a major microbe-associated molecular pattern (MAMP) of Gram-positive bacteria. However, many detai...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-161
更新日期:2012-12-15 00:00:00
abstract:BACKGROUND:Ganoderic acids (GAs), derived from the medicinal mushroom Ganoderma lucidum, possess anticancer and other important pharmacological activities. To improve production of GAs, a homologous farnesyl diphosphate synthase (FPS) gene was overexpressed in G. lucidum. Moreover, the influence of FPS gene overexpress...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1164-3
更新日期:2019-06-28 00:00:00
abstract:BACKGROUND:The biocontrol strain Pseudomonas chlororaphis GP72 isolated from the green pepper rhizosphere synthesizes three antifungal phenazine compounds, 2-Hydroxyphenazine (2-OH-PHZ), 2-hydroxy-phenazine-1-carboxylic acid (2-OH-PCA) and phenazine-1-carboxylic acid (PCA). PCA has been a commercialized antifungal pest...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0529-0
更新日期:2016-07-28 00:00:00
abstract:BACKGROUND:A stereoisomer of inositol, scyllo-inositol (SI), has been regarded as a promising therapeutic agent for Alzheimer's disease. However, this compound is relatively rare, whereas another stereoisomer of inositol, myo-inositol (MI) is abundant in nature. Bacillus subtilis 168 has the ability to metabolize inosi...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0682-0
更新日期:2017-04-21 00:00:00
abstract::Amylase plays an important role in biotechnology industries, and Gram-positive bacterium Bacillus subtilis is a major host to produce heterogeneous α-amylases. However, the secretion stress limits the high yield of α-amylase in B. subtilis although huge efforts have been made to address this secretion bottleneck. In t...
journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/s12934-017-0738-1
更新日期:2017-07-19 00:00:00
abstract:BACKGROUND:L-Alanyl-L-glutamine (AQ) is a functional dipeptide with high water solubility, good thermal stability and high bioavailability. It is widely used in clinical treatment, post-operative rehabilitation, sports health care and other fields. AQ is mainly produced via chemical synthesis which is complicated, time...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01369-2
更新日期:2020-06-11 00:00:00
abstract:BACKGROUND:Despite the progressive miniaturization of bioreactors for screening purposes, shake flasks are still widespread in biotechnological laboratories and industry as cultivation vessels. Shake flasks are often applied as the first or second step in applications such as strain screening or media optimization. Thu...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0444-4
更新日期:2016-02-24 00:00:00
abstract:BACKGROUND:High-throughput screening methods assume that the output measured is representative of changes in metabolic flux toward the desired product and is not affected by secondary phenotypes. However, metabolic engineering can result in unintended phenotypes that may go unnoticed in initial screening. The red pigme...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0381-7
更新日期:2015-11-26 00:00:00
abstract:BACKGROUND:The methylotrophic yeast Pichia pastoris is a well-studied host organism for recombinant protein production, which is usually regulated either by a constitutive promoter (e.g. promoter of glyceraldehyde-3-phosphate dehydrogenase; PGAP) or an inducible promoter (e.g. promoter of alcohol oxidase 1; PAOX1). Bot...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0768-8
更新日期:2017-09-13 00:00:00
abstract:BACKGROUND:C4-dicarboxylic acids, including malic acid, fumaric acid and succinic acid, are valuable organic acids that can be produced and secreted by a number of microorganisms. Previous studies on organic acid production by Aspergillus carbonarius, which is capable of producing high amounts of citric acid from varie...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0660-6
更新日期:2017-03-14 00:00:00
abstract:BACKGROUND:Plasmid expression is a popular method in studies of MVA pathway for isoprenoid production in Escherichia coli. However, heterologous gene expression with plasmid is often not stable and might burden growth of host cells, decreases cell mass and product yield. In this study, MVA pathway was divided into thre...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0607-3
更新日期:2016-12-01 00:00:00
abstract:BACKGROUND:The deep-sea environment harbors a vast pool of novel enzymes. Owing to the limitations of cultivation, cultivation-independent has become an effective method for mining novel enzymes from the environment. Based on a deep-sea sediment metagenomics library, lipolytic-positive clones were obtained by activity-...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0864-4
更新日期:2018-01-30 00:00:00
abstract:BACKGROUND:We evaluated the functional capacity of plantaricin-producing Lactobacillus plantarum SF9C and S-layer-carrying Lactobacillus brevis SF9B to withstand gastrointestinal transit and to compete among the gut microbiota in vivo. Considering the probiotic potential of Lb. brevis SF9B, this study aims to investiga...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01365-6
更新日期:2020-05-19 00:00:00
abstract:BACKGROUND:Mannosylglycerate (MG) is one of the most widespread compatible solutes among marine microorganisms adapted to hot environments. This ionic solute holds excellent ability to protect proteins against thermal denaturation, hence a large number of biotechnological and clinical applications have been put forward...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-1023-7
更新日期:2018-11-16 00:00:00
abstract:BACKGROUND:Acetic acid bacteria (AAB) are widely applied in food, bioengineering and medicine fields. However, the acid stress at low pH conditions limits acetic acid fermentation efficiency and high concentration of vinegar production with AAB. Therefore, how to enhance resistance ability of the AAB remains as the maj...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0717-6
更新日期:2017-06-15 00:00:00
abstract:BACKGROUND:Oxidized glutathione (GSSG) is the preferred form for industrial mass production of glutathione due to its high stability compared with reduced glutathione (GSH). In our previous study, over-expression of the mitochondrial thiol oxidase ERV1 gene was the most effective for high GSSG production in Saccharomyc...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0658-0
更新日期:2017-03-15 00:00:00
abstract:BACKGROUND:Cell surface display of recombinant proteins has become a powerful tool for biotechnology and biomedical applications. As a model eukaryotic microorganism, Saccharomyces cerevisiae is an ideal candidate for surface display of heterologous proteins. However, the frequently used commercial yeast surface displa...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1133-x
更新日期:2019-05-18 00:00:00
abstract:BACKGROUND:Microalgae has been concerned as a potential source of biodiesel in the recent years. However, it is costly to harvest microalgae as it is commonly cultured in water and the cells are too small to harvest. In order to reduce the cost of cultivation and harvesting, it is important to improve the biomass produ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0142-z
更新日期:2014-10-01 00:00:00
abstract:BACKGROUND:Heterologous expression of biosynthetic gene clusters of natural microbial products has become an essential strategy for titer improvement and pathway engineering of various potentially-valuable natural products. A Streptomyces artificial chromosomal conjugation vector, pSBAC, was previously successfully app...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0708-7
更新日期:2017-05-31 00:00:00
abstract:BACKGROUND:Co-expression of two distinct guide RNAs (gRNAs) has been used to facilitate the application of CRISPR/Cas9 system in fields such as large genomic deletion. The paired gRNAs are often placed adjacently in the same direction and expressed individually by two identical promoters, constituting direct repeats (D...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01321-4
更新日期:2020-03-10 00:00:00
abstract::Innovative new genome engineering technologies for manipulating chromosomes have appeared in the last decade. One of these technologies, recombination mediated genetic engineering (recombineering) allows for precision DNA engineering of chromosomes and plasmids in Escherichia coli. Single-stranded DNA recombineering (...
journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/1475-2859-13-S1-S10
更新日期:2014-08-29 00:00:00
abstract:BACKGROUND:The sesquiterpenoid abscisic acid (ABA) is mostly known for regulating developmental processes and abiotic stress responses in higher plants. Recent studies show that ABA also exhibits a variety of pharmacological activities. Affordable and sustainable production will be required to utilize the compound in a...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1257-z
更新日期:2019-11-25 00:00:00
abstract:BACKGROUND:The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals. Itaconic acid (IA) has ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0527-2
更新日期:2016-07-28 00:00:00
abstract:BACKGROUND:In Escherichia coli many heterologous proteins are produced in the periplasm. To direct these proteins to the periplasm, they are equipped with an N-terminal signal sequence so that they can traverse the cytoplasmic membrane via the protein-conducting Sec-translocon. For poorly understood reasons, the produc...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-24
更新日期:2013-03-12 00:00:00
abstract:BACKGROUND:Sesquiterpenes are a class of natural products with a diverse range of attractive industrial proprieties. Due to economic difficulties of sesquiterpene production via extraction from plants or chemical synthesis there is interest in developing alternative and cost efficient bioprocesses. The hydrocarbon α-sa...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-117
更新日期:2012-08-31 00:00:00
abstract:BACKGROUND:Terpenoids constitute a large family of natural products, attracting commercial interest for a variety of uses as flavours, fragrances, drugs and alternative fuels. Saccharomyces cerevisiae offers a versatile cell factory, as the precursors of terpenoid biosynthesis are naturally synthesized by the sterol bi...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-4
更新日期:2011-01-28 00:00:00
abstract:BACKGROUND:Nowadays, microbial infections have caused increasing economic losses in aquaculture industry and deteriorated worldwide environments. Many of these infections are caused by opportunistic pathogens through cell-density mediated quorum sensing (QS). The disruption of QS, known as quorum quenching (QQ), is an ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01448-4
更新日期:2020-10-07 00:00:00