Abstract:
BACKGROUND:Proteins can be secreted from a host organism with the aid of N-terminal secretion signals. The budding yeast Pichia pastoris (Komagataella sp.) is widely employed to secrete proteins of academic and industrial interest. For this yeast, the most commonly used secretion signal is the N-terminal portion of pre-pro-α-factor from Saccharomyces cerevisiae. However, this secretion signal promotes posttranslational translocation into the endoplasmic reticulum (ER), so proteins that can fold in the cytosol may be inefficiently translocated and thus poorly secreted. In addition, if a protein self-associates, the α-factor pro region can potentially cause aggregation, thereby hampering export from the ER. This study addresses both limitations of the pre-pro-α-factor secretion signal. RESULTS:We engineered a hybrid secretion signal consisting of the S. cerevisiae Ost1 signal sequence, which promotes cotranslational translocation into the ER, followed by the α-factor pro region. Secretion and intracellular localization were assessed using as a model protein the tetrameric red fluorescent protein E2-Crimson. When paired with the α-factor pro region, the Ost1 signal sequence yielded much more efficient secretion than the α-factor signal sequence. Moreover, an allelic variant of the α-factor pro region reduced aggregation of the E2-Crimson construct in the ER. The resulting improved secretion signal enhanced secretion of E2-Crimson up to 20-fold compared to the levels obtained with the original α-factor secretion signal. Similar findings were obtained with the lipase BTL2, which exhibited 10-fold enhanced secretion with the improved secretion signal. CONCLUSIONS:The improved secretion signal confers dramatic benefits for the secretion of certain proteins from P. pastoris. These benefits are likely to be most evident for proteins that can fold in the cytosol and for oligomeric proteins.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Barrero JJ,Casler JC,Valero F,Ferrer P,Glick BSdoi
10.1186/s12934-018-1009-5subject
Has Abstractpub_date
2018-10-12 00:00:00pages
161issue
1issn
1475-2859pii
10.1186/s12934-018-1009-5journal_volume
17pub_type
杂志文章abstract:BACKGROUND:Recombinant antibodies can be produced in different formats and different expression systems. Single chain variable fragments (scFvs) represent an attractive alternative to full-length antibodies and they can be easily produced in bacteria or yeast. However, the scFvs exhibit monovalent antigen-binding prope...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-109
更新日期:2011-12-15 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: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:Erythritol is a four-carbon sugar alcohol with sweetening properties that is used by the agro-food industry as a food additive. In the yeast Yarrowia lipolytica, the last step of erythritol synthesis involves the reduction of erythrose by specific erythrose reductase(s). In the earlier report, an erythrose r...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0982-z
更新日期:2018-08-29 00:00:00
abstract::Like natural viruses, manmade protein cages for drug delivery are to be ideally formed by repetitive subunits with self-assembling properties, mimicking viral functions and molecular organization. Naturally formed nanostructures (such as viruses, flagella or simpler protein oligomers) can be engineered to acquire spec...
journal_title:Microbial cell factories
pub_type: 社论
doi:10.1186/1475-2859-9-101
更新日期:2010-12-30 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:Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metaboli...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01466-2
更新日期:2020-11-23 00:00:00
abstract:BACKGROUND:Lactobacillus ruminis is a poorly characterized member of the Lactobacillus salivarius clade that is part of the intestinal microbiota of pigs, humans and other mammals. Its variable abundance in human and animals may be linked to historical changes over time and geographical differences in dietary intake of...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-S1-S12
更新日期:2011-08-30 00:00:00
abstract:BACKGROUND:The application of chemical dispersants as a response to marine oil spills is raising concerns related to their potential toxicity also towards microbes involved in oil biodegradation. Hence, oil spills occurring under marine environments necessitate the application of biodispersants that are highly active, ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0797-3
更新日期:2017-11-02 00:00:00
abstract::Nanotechnology is increasingly using both materials and nano-objects synthesized by living beings, most of them produced by microbial cells. Emerging technologies and highly integrative approaches (such as 'omics and systems biology), that have been largely proven successful for the production of proteins and secondar...
journal_title:Microbial cell factories
pub_type: 社论
doi:10.1186/1475-2859-9-53
更新日期:2010-07-05 00:00:00
abstract:BACKGROUND:A number of valuable candidates as tuberculosis vaccine have been reported, some of which have already entered clinical trials. The new vaccines, especially subunit vaccines, need multiple administrations in order to maintain adequate life-long immune memory: this demands for high production levels and degre...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-115
更新日期:2013-11-19 00:00:00
abstract:BACKGROUND:In past years research has focused on the development of alternative Gram positive bacterial expression systems to produce industrially relevant proteins. Brevibacillus choshinensis is an easy to handle non-sporulating bacterium, lacking extracellular proteases, that has been already shown to provide a high ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-12
更新日期:2013-02-01 00:00:00
abstract:BACKGROUND:Gene therapy has gained an increasing interest in its anti-tumor efficiency. However, numerous efforts are required to promote them to clinics. In this study, a novel and efficient delivery platform based on bacterial magnetosomes (BMs) were developed, and the efficiency of BMs in delivering small interferin...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0830-6
更新日期:2017-11-28 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:Calreticulin (CRT) resides in the endoplasmic reticulum (ER) and functions to chaperone proteins, ensuring proper folding, and intracellular Ca(2+) homeostasis. Emerging evidence shows that CRT is a multifunctional protein with significant roles in physiological and pathological processes with presence both ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0356-8
更新日期:2015-10-15 00:00:00
abstract:BACKGROUND:Understanding the metabolic mechanism of sterols to produce valuable steroid intermediates in mycobacterium by a noncoding small RNA (sRNA) view is still limited. In the work, RNA-seq was implemented to investigate the noncoding transcriptome of Mycobacterium neoaurum (Mn) in the transformation process of st...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0462-2
更新日期:2016-04-25 00:00:00
abstract:BACKGROUND:Oxygen-evolving photoautotrophic organisms, like cyanobacteria, protect their photosynthetic machinery by a number of regulatory mechanisms, including alternative electron transfer pathways. Despite the importance in modulating the electron flux distribution between the photosystems, alternative electron tra...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1238-2
更新日期:2019-11-05 00:00:00
abstract:BACKGROUND:Due to their high energy density and compatible physical properties, several monoterpenes have been investigated as potential renewable transportation fuels, either as blendstocks with petroleum or as drop-in replacements for use in vehicles (both heavy and light-weight) or in aviation. Sustainable microbial...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1099-8
更新日期:2019-03-18 00:00:00
abstract:BACKGROUND:Pichia pastoris is one of the most important host organisms for the recombinant production of proteins in industrial biotechnology. To date, strain specific parameters, which are needed to set up feeding profiles for fed batch cultivations, are determined by time-consuming continuous cultures or consecutive ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-14
更新日期:2011-03-03 00:00:00
abstract:BACKGROUND:Corynebacterium glutamicum was used as a metabolic engineering chassis for production of crude violacein (mixture of violacein and deoxyviolacein) due to Corynebacterium's GRAS status and advantages in tryptophan fermentation. The violacein is a commercially potential compound with various applications deriv...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0545-0
更新日期:2016-08-24 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:BACKGROUND:Microbial genome editing is a powerful tool to modify chromosome in way of deletion, insertion or replacement, which is one of the most important techniques in metabolic engineering research. The emergence of CRISPR/Cas9 technique inspires various genomic editing methods. RESULTS:In this research, the goal ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0605-5
更新日期:2016-12-01 00:00:00
abstract:BACKGROUND:Hepatitis C virus (HCV) is a common and leading cause for liver cirrhosis and hepatocellular carcinoma. Current therapies to treat HCV infection are shown to be partially effective and poorly tolerated. Therefore, ample efforts are underway to rationally design therapies targeting the HCV non-structural prot...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-111
更新日期:2013-11-16 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:Spore-forming bacteria of the Bacillus genus are widely used probiotics known to exert their beneficial effects also through the stimulation of the host immune response. The oral delivery of B. toyonensis spores has been shown to improve the immune response to a parenterally administered viral antigen in mic...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01308-1
更新日期:2020-02-19 00:00:00
abstract:BACKGROUND:Rhodosporidium and Rhodotorula are two genera of oleaginous red yeast with great potential for industrial biotechnology. To date, there is no effective method for inducible expression of proteins and RNAs in these hosts. RESULTS:We have developed a luciferase gene reporter assay based on a new codon-optimiz...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0357-7
更新日期:2015-10-26 00:00:00
abstract:BACKGROUND:Our laboratory has constructed a Bacillus stearothermophilus α-amylase (AmyS) derivative with excellent enzymatic properties. Bacillus subtilis is generally regarded as safe and has excellent protein secretory capability, but heterologous extracellular production level of B. stearothermophilus α-amylase in B...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1119-8
更新日期:2019-04-11 00:00:00
abstract:BACKGROUND:Mucosal delivery of therapeutic proteins using genetically modified strains of lactic acid bacteria (gmLAB) is being investigated as a new therapeutic strategy. METHODS:We developed a strain of gmLAB, Lactococcus lactis NZ9000 (NZ-HO), which secretes the anti-inflammatory molecule recombinant mouse heme oxy...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0378-2
更新日期:2015-11-25 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:Intracellular metabolism of glucocorticoid hormones plays an important role in the pathogenesis of metabolic syndrome and regulates, among many physiological processes, collagen metabolism in skin. At the peripheral level the concentration of active glucocorticoids is mainly regulated by the 11β-hydroxystero...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0450-6
更新日期:2016-03-15 00:00:00