Abstract:
:Pichia pastoris is widely used as a host system for heterologous protein expression in both academia and industry. Production is typically accomplished by a fed-batch induction process that is known to have negative impacts on cell physiology that impose limits on both protein yields and quality. We have analysed recombinant protein production in chemostat cultures to understand the physiological responses associated with methanol-induced production of two human lysozyme variants with different degrees of misfolding by P. pastoris. Confounding variables associated with nutrient stress or growth-rate are minimised during steady-state growth in chemostats. Comparison of transcriptome-level data obtained during the non-inducing and inducing steady states identified changes in expression of only about 1 % of the genome during production of either an amyloidogenic human lysozyme variant prone to intracellular aggregation (I56T) or a misfolded but secretable variant (T70N), indicating near-complete acclimation to their production. A marked, but temporary, stress response involving both the unfolded protein response (UPR) and ER-associated degradation pathway was observed during the transient between steady states, particularly following induction of the T70N variant synthesis, and was accompanied by changes in expression of around 50 antisense transcripts. The results suggest that optimal heterologous protein production could best be achieved by a continuous process that minimises the number of methanol-induced transients experienced by the cultures. The processing of HAC1 mRNA required for the UPR was found to be constitutive in the culture conditions used, even in the absence of recombinant protein induction.
journal_name
Appl Microbiol Biotechnoljournal_title
Applied microbiology and biotechnologyauthors
Hesketh AR,Castrillo JI,Sawyer T,Archer DB,Oliver SGdoi
10.1007/s00253-013-5186-1subject
Has Abstractpub_date
2013-11-01 00:00:00pages
9747-9762issue
22eissn
0175-7598issn
1432-0614journal_volume
97pub_type
杂志文章abstract::Aerobic granules are cultivated by a single bacterial strain, Acinetobacter calcoaceticus, in a sequencing batch reactor (SBR). This strain presents as a good phenol reducer and an efficient auto coagulator in the presence of phenol, mediated by heat-sensitive adhesins proteins. Stable 2.3-mm granules were formed in t...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-007-1325-x
更新日期:2008-03-01 00:00:00
abstract::This study investigated the effects of vanillin on methanogenesis and rumen fermentation, and the responses of ruminal protein-degrading bacteria to vanillin (at concentrations of 0, 0.76 and 1.52 g/L), essential oils (clove oil, 1 g/L; origanum oil, 0.50 g/L, and peppermint oil, 1 g/L), and quillaja saponin (at conce...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-013-4930-x
更新日期:2014-01-01 00:00:00
abstract::Biodegradation of petroleum compounds in saline environments seems intricate and needs more attention. In this study, tetracosane was used to enrich alkane-degrading bacteria from oil-contaminated saline soils. Among the isolates, strain Qtet3, with the highest 16s rRNA gene sequence similarity to Alcanivorax dieselol...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-010-3049-6
更新日期:2011-04-01 00:00:00
abstract::The white button mushroom Agaricus bisporus is economically the most important commercially produced edible fungus. It is grown on carbon- and nitrogen-rich substrates, such as composted cereal straw and animal manure. The commercial mushroom production process is usually performed in buildings or tunnels under highly...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-017-8294-5
更新日期:2017-06-01 00:00:00
abstract::Yarrowia lipolytica is an industrial yeast that has been used in the sustainable production of fatty acid-derived and lipid compounds due to its high growth capacity, genetic tractability, and oleaginous properties. This investigation examines the possibility of utilizing urea or urine as an alternative to ammonium su...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-8769-z
更新日期:2018-03-01 00:00:00
abstract::Catechin is a plant polyphenol composed of epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG), and epigallocatechin gallate (EGCG) as diastereoisomers. Among the various classes of flavonoids, catechin was found to be the most powerful free radical scavenger, scavenging the reactive oxygen species (RO...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-016-7492-x
更新日期:2016-07-01 00:00:00
abstract::α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-013-5145-x
更新日期:2014-03-01 00:00:00
abstract::To develop microbial production method for prenyl alcohols (e.g., (E,E)-farnesol (FOH), (E)-nerolidol (NOH), and (E,E,E)-geranylgeraniol (GGOH)), the genes encoding enzymes in the mevalonate and prenyl diphosphate pathways were overexpressed in Saccharomyces cerevisiae, and the resultant transformants were evaluated a...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-008-1807-5
更新日期:2009-04-01 00:00:00
abstract::The manganese superoxide dismutase gene (sodM) is very highly expressed in Aspergillus oryzae. To elucidate the basis for this high-level expression, deletion analysis of the promoter was undertaken using beta-glucuronidase (GUS) as a reporter. Deletion of a 63-bp sequence from -200 to -138 in the 1,038-bp sodM promot...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-006-0388-4
更新日期:2006-10-01 00:00:00
abstract::β-Glucuronidase is a lysosomal enzyme and a molecular model of a class of therapeutics approved as enzyme replacement therapies for lysosomal storage diseases. Understanding the effect of bioreactor process variables on the production and quality of the biologics is critical for maintaining quality and efficacy of the...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-019-09889-7
更新日期:2019-08-01 00:00:00
abstract::The microbial oxidation of ebastine to carebastine was investigated. Among the 15 micro-organisms examined, only the Cunninghamella strains showed the desired biotransformation. Cunninghamella blakesleeana oxidised the substrate within 7 days, via the intermediates alcohol and aldehyde, mainly to carebastine, the corr...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/BF00178610
更新日期:1996-02-01 00:00:00
abstract::The impact of glucose on glycerol metabolism, especially on 3-hydroxypropionaldehyde (3-HPA) accumulation by resting cells of Lactobacillus reuteri has been investigated. Two systems were used in the study: MRS(-) (modified MRS - omitting glucose, acetate and Tween 80) and distilled water (H(2)O). In MRS(-), addition ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-002-1002-z
更新日期:2002-07-01 00:00:00
abstract::The efficient production of recombinant proteins such as antibodies typically involves the screening of an extravagant number of clones in order to finally select a stable and high-producing cell line. Thereby, the underlying principles of a powerful protein machinery, but also potential expression limitations, often ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-5584-z
更新日期:2014-07-01 00:00:00
abstract::Gluconic acid secretion mediated by the direct oxidation of glucose by pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenase (GDH) is responsible for mineral phosphate solubilization in Gram-negative bacteria. Herbaspirillum seropedicae Z67 (ATCC 35892) genome encodes GDH apoprotein but lacks genes for the bi...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-5610-1
更新日期:2014-06-01 00:00:00
abstract::Pseudomonas chlororaphis is a plant-associated bacterium with reported antagonistic activity against different organisms and plant growth-promoting properties. P. chlororaphis possesses exciting biotechnological features shared with another Pseudomonas with a nonpathogenic phenotype. Part of the antagonistic role of P...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-020-10913-4
更新日期:2020-12-01 00:00:00
abstract::Novel alginate hydrogels with silver nanoparticles (AgNPs) and honey components were produced with the aim to target multidrug-resistant bacterial strains causing nosocomial wound infections. AgNP synthesis was optimized in highly concentrated honey solutions so that a 5-month stable, colloid solution with 50% of hone...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-019-10055-2
更新日期:2019-10-01 00:00:00
abstract::Insertion specificity of mobile genetic elements is a rather complex aspect of DNA transposition, which, despite much progress towards its elucidation, still remains incompletely understood. We report here the results of a meta-analysis of IS2 target sites from genomic, phage, and plasmid DNA and find that newly acqui...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-5695-6
更新日期:2014-08-01 00:00:00
abstract::Bacillus thuringiensis bacteria show insecticidal activities that rely upon the production of insecticidal crystal proteins, which are encoded by cry or cyt genes and can target a variety of insect pests. It has been shown that cry1Ac is the only cry gene in B. thuringiensis subsp. kurstaki HD73 (B. thuringiensis HD73...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-8836-5
更新日期:2018-04-01 00:00:00
abstract::Photosynthetic microorganisms produce relatively large amounts of physiologically active materials which stimulate the physiological activity of other organisms. In this study, mammalian HeLa cells were cultured in different culture media which were Dulbecco's modified Eagle medium (DMEM) with newborn calf serum (NCS)...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-012-4653-4
更新日期:2013-11-01 00:00:00
abstract::Regulatable promoters are important genetic tools, particularly for assigning function to essential and redundant genes. They can also be used to control the expression of enzymes that influence metabolic flux or protein secretion, thereby optimizing product yield in bioindustry. This review will focus on regulatable ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-018-8887-7
更新日期:2018-05-01 00:00:00
abstract::Sialylated oligosaccharides are known to have beneficial effects, such as increasing the level of bifidobacteria, reducing the levels of blood endotoxin and blood ammonia, and enhancing the body's immune system. However, it is unknown whether sialylated lactuloses have modulatory effects on the intestinal microbiota. ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-019-10169-7
更新日期:2019-11-01 00:00:00
abstract::Microbial transglutaminase (MTG) from Streptomyces mobaraensis has been widely used for crosslinking proteins in order to acquire products with improved properties. To improve the yield and enable a facile and efficient purification process, recombinant vectors, harboring various heterologous signal peptide-encoding f...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-018-9000-y
更新日期:2018-07-01 00:00:00
abstract::Immobilization of the Geotrichum candidum (CCT 1205) cell with functionalized silica creates promising biocatalysts for production of ɛ-caprolactone. The results obtained by immobilization of the whole cell on SiO2-NH2 and SiO2-SH supports indicate that the presence of reactive functional groups on the support may pro...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-020-10875-7
更新日期:2020-10-01 00:00:00
abstract::Aspartic proteases exhibit optimum enzyme activity under acidic conditions and have been extensively used in food, fermentation, and leather industries. In this study, a novel aspartic protease precursor (proTlAPA1) from Talaromyces leycettanus was identified and successfully expressed in Pichia pastoris. Subsequently...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-020-10569-0
更新日期:2020-06-01 00:00:00
abstract::Paddy fields are a significant source of methane and contribute up to 20% of total methane emissions from wetland ecosystems. These inundated, anoxic soils featuring abundant nitrogen compounds and methane are an ideal niche for nitrate-dependent anaerobic methanotrophs. After 2 years of enrichment with a continuous s...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-017-8416-0
更新日期:2017-09-01 00:00:00
abstract::We report on a pheromone-based inter-species communication system, allowing for a controlled cell-cell communication between the two species Saccharomyces cerevisiae and Schizosaccharomyces pombe as a proof of principle. It exploits the mating response pathways of the two yeast species employing the pheromones, α- or ...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s00253-014-6133-5
更新日期:2015-02-01 00:00:00
abstract::Pimaricin (natamycin) is a small polyene macrolide antibiotic used worldwide. This efficient antimycotic and antiprotozoal agent, produced by several soil bacterial species of the genus Streptomyces, has found application in human therapy, in the food and beverage industries and as pesticide. It displays a broad spect...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-015-7077-0
更新日期:2016-01-01 00:00:00
abstract::Crude peroxidase preparations from the lignocellulose-degrading actinomycete, Streptomyces viridosporus T7A, were shown to decolorize several azo dye isomers and showed a correlation of dye structure to degradability similar to that shown by fungal Mn-peroxidase, an enzyme not previously described in actinomycetes. Ad...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051208
更新日期:1998-05-01 00:00:00
abstract::Plant-parasitic nematodes are estimated to cause global annual losses of more than US$ 100 billion. The number of registered nematicides has declined substantially over the last 25 years due to concerns about their non-specific mechanisms of action and hence their potential toxicity and likelihood to cause environment...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s00253-015-7233-6
更新日期:2016-05-01 00:00:00
abstract::A general procedure for the high yield immobilization of enzymes with the help of specific anti-enzyme antibodies is described. Polyclonal antibodies were raised against Aspergillus niger glucose oxidase and horseradish peroxidase in rabbits and the gamma globulin (IgG) fraction from the immune sera isolated by ammoni...
journal_title:Applied microbiology and biotechnology
pub_type: 杂志文章
doi:10.1007/s002530051534
更新日期:1999-09-01 00:00:00