Construction of second generation protease-deficient hosts of Bacillus subtilis for secretion of foreign proteins.

Abstract:

:Although one of the major factors limiting the application of Bacillus subtilis as an expression host has been its production of at least eight extracellular proteases, researchers have also noticed that some proteases benefited the secretion of foreign proteins at times. Therefore, to maximize the yield of a foreign protein, the proteases should be selectively inactivated. This raises a new question that how to identify the favorable and unfavorable proteases for a target protein. Here, an evaluation system containing nine mutant strains of B. subtilis 168 was developed to address this question. The mutant strain PD8 has all the eight proteases inactivated whereas each of the other eight mutant strains expresses only one kind of these eight proteases. The target protein is secreted in these nine mutant strains; if the production of target protein in a mutant strain is higher than that in strain PD8, the corresponding protease is regarded as favorable. Accordingly, the optimal protease-deficient host is constructed through inactivating the unfavorable proteases. The effectiveness of this system was confirmed by expressing three foreign proteins. This study provides a strategy for improving the secretion of a foreign protein in B. subtilis through tailoring a personalized protease-deficient host.

journal_name

Biotechnol Bioeng

authors

Zhao L,Ye B,Zhang Q,Cheng D,Zhou C,Cheng S,Yan X

doi

10.1002/bit.26992

subject

Has Abstract

pub_date

2019-08-01 00:00:00

pages

2052-2060

issue

8

eissn

0006-3592

issn

1097-0290

journal_volume

116

pub_type

杂志文章
  • Autoradiographic determination of mass-transfer limitations in immobilized cell reactors.

    abstract::Pseudomonas putida cells were grown in confined volumes in dual-membrane immobilized cell reactors constructed from microporous polyethylene hollow fibers and silicone rubber tubules as a model system for the study of mass transport in microbial aggregates. Local cell concentrations in the reactors reached 300 g dry m...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260340307

    authors: Karel SF,Robertson CR

    更新日期:1989-07-01 00:00:00

  • Harvesting of Scenedesmus obliquus in wastewaters: auto- or bioflocculation?

    abstract::Autoflocculation and bioflocculation are considered to be the most promising means for the economical harvesting of microalgae. We have therefore studied these phenomena with cultures of Scenedesmus obliquus produced during biological tertiary wastewater treatment. The quantity of extracellular polymers produced durin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260300707

    authors: Lavoie A,de la Noüe J

    更新日期:1987-11-01 00:00:00

  • Characterization of extremely thermostable enzymatic breakers (alpha-1,6-galactosidase and beta-1,4-mannanase) from the hyperthermophilic bacterium Thermotoga neapolitana 5068 for hydrolysis of guar gum.

    abstract::An alpha-galactosidase and a beta-mannanase produced by the hyperthermophilic bacterium, Thermotoga neapolitana 5068 (TN5068), separately and together, were evaluated for their ability to hydrolyze guar gum in relation to viscosity reduction of guar-based hydraulic fracturing fluids used in oil and gas well stimulatio...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(SICI)1097-0290(19961020)52:2<332::AID-BIT

    authors: McCutchen CM,Duffaud GD,Leduc P,Petersen AR,Tayal A,Khan SA,Kelly RM

    更新日期:1996-10-20 00:00:00

  • Active-site titration of serine proteases in organic solvents.

    abstract::Calculation of kinetic constants of an enzymatic reaction in organic solvents requires knowledge of the functional active-site concentration in organic solvents, and this can be significantly different than that in water. An experimental method for active-site titration of serine proteases in organic media has been de...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(SICI)1097-0290(19960505)50:3<329::AID-BIT

    authors: Wangikar PP,Carmichael D,Clark DS,Dordick JS

    更新日期:1996-05-05 00:00:00

  • Effect of cell lysates on retroviral transduction efficiency of cells in suspension culture.

    abstract::Recombinant retroviruses are effective vectors able to integrate transgenes into the target cell's genome to achieve longer-term expression. This study investigates the effect of cell lysis products, a common cell culture by-product, on the transduction of suspension cells by gammaretroviral vectors. Cell lysates deri...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22634

    authors: Beauchesne PR,Bruce KJ,Bowen BD,Piret JM

    更新日期:2010-04-15 00:00:00

  • Targeted gene delivery to CD117-expressing cells in vivo with lentiviral vectors co-displaying stem cell factor and a fusogenic molecule.

    abstract::The development of a lentiviral system to deliver genes to specific cell types could improve the safety and the efficacy of gene delivery. Previously, we have developed an efficient method to target lentivectors to specific cells via an antibody-antigen interaction in vitro and in vivo. We report herein a targeted len...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22378

    authors: Froelich S,Ziegler L,Stroup K,Wang P

    更新日期:2009-09-01 00:00:00

  • Extracellular hemoglobin combined with an O2 -generating material overcomes O2 limitation in the bioartificial pancreas.

    abstract::The bioartificial pancreas encapsulating pancreatic islets in immunoprotective hydrogel is a promising therapy for Type 1 diabetes. As pancreatic islets are highly metabolically active and exquisitely sensitive to hypoxia, maintaining O2 supply after transplantation remains a major challenge. In this study, we address...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26913

    authors: Mouré A,Bacou E,Bosch S,Jegou D,Salama A,Riochet D,Gauthier O,Blancho G,Soulillou JP,Poncelet D,Olmos E,Bach JM,Mosser M

    更新日期:2019-05-01 00:00:00

  • Towards the industrialization of new biosurfactants: Biotechnological opportunities for the lactone esterase gene from Starmerella bombicola.

    abstract::Although sophorolipids (SLs) produced by S. bombicola are a real showcase for the industrialization of microbial biosurfactants, some important drawbacks are associated with this efficient biological process, e.g., the simultaneous production of acidic and lactonic SLs. Depending on the application, there is a require...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25815

    authors: Roelants SL,Ciesielska K,De Maeseneire SL,Moens H,Everaert B,Verweire S,Denon Q,Vanlerberghe B,Van Bogaert IN,Van der Meeren P,Devreese B,Soetaert W

    更新日期:2016-03-01 00:00:00

  • Biofilm parameters influencing biocide efficacy.

    abstract::The influence of biofilm areal cell density, species composition, and the presence of abiotic particles on the disinfection and removal of bacterial biofilms by monochloramine was investigated. Mono- and binary population biofilms of Pseudomonas aeruginosa and Klebsiella pneumoniae were grown on stainless-steel slides...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260460608

    authors: Srinivasan R,Stewart PS,Griebe T,Chen CI,Xu X

    更新日期:1995-06-20 00:00:00

  • Mitigation of ammonia inhibition by internal dilution in high-rate anaerobic digestion of food waste leachate and evidences of microbial community response.

    abstract::A high-rate anaerobic digestion of food waste leachate were tested using intermittent continuously stirred tank reactors (iCSTRs) to evaluate how severe ammonia inhibition could be mitigated with internal dilution strategy, and to identify how bacterial and archaeal community respond in genus and species level. Experi...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25968

    authors: Yun YM,Kim DH,Cho SK,Shin HS,Jung KW,Kim HW

    更新日期:2016-09-01 00:00:00

  • Analysis of a continuous, aerobic, fixed-film bioreactor. I. Steady-state behavior.

    abstract::The analysis of a continuous, aerobic, fixed-film bioreactor is performed by simulating the behavior of penicillin production in a three-phase fluidized bed. Rigorous mathematical models are developed for a fluidized-bed fermentor in which bioparticles are fluidized by the liquid medium and air. The steady-state perfo...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260260509

    authors: Park Y,Davis ME,Wallis DA

    更新日期:1984-05-01 00:00:00

  • Electric field control of lipase membrane activity.

    abstract::Lipase (EC 3.1.1.3., from Pseudomonas sp.) was entrapped in collagen membrane containing liquid crystal (4-methoxybenzilidene-4'-n-butylaniline). The activity of the lipase-liquid crystal membrane at an applied voltage of 4 V was 3.4 compared to a membrane tested without imposition of an external electric field. A lin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260191007

    authors: Karube I,Yugeta Y,Suzuki S

    更新日期:1977-10-01 00:00:00

  • Computing the various pathways of penicillin synthesis and their molar yields.

    abstract::More than 80 years after its discovery, penicillin is still a widely used and commercially highly important antibiotic. Here, we analyse the metabolic network of penicillin synthesis in Penicillium chrysogenum based on the concept of elementary flux modes. In particular, we consider the synthesis of the invariant mole...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25694

    authors: Prauße MT,Schäuble S,Guthke R,Schuster S

    更新日期:2016-01-01 00:00:00

  • A novel sugar analog enhances sialic acid production and biotherapeutic sialylation in CHO cells.

    abstract::A desirable feature of many therapeutic glycoprotein production processes is to maximize the final sialic acid content. In this study, the effect of applying a novel chemical analog of the sialic acid precursor N-acetylmannosamine (ManNAc) on the sialic acid content of cellular proteins and a model recombinant glycopr...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26291

    authors: Yin B,Wang Q,Chung CY,Bhattacharya R,Ren X,Tang J,Yarema KJ,Betenbaugh MJ

    更新日期:2017-08-01 00:00:00

  • Effect of the applied organic load rate on biodegradable polymer production by mixed microbial cultures in a sequencing batch reactor.

    abstract::This article studies the operation of a new process for the production of biopolymers (polyhydroxyalkanoates, PHAs) at different applied organic load rates (OLRs). The process is based on the aerobic enrichment of activated sludge to obtain mixed cultures able to store PHAs at high rates and yields. A mixture of aceti...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20683

    authors: Dionisi D,Majone M,Vallini G,Di Gregorio S,Beccari M

    更新日期:2006-01-05 00:00:00

  • Alginate microparticles as novel carrier for oral insulin delivery.

    abstract::Alginate microparticles produced by emulsification/internal gelation were investigated as a promising carrier for insulin delivery. The procedure involves the dispersion of alginate solution containing insulin protein, into a water immiscible phase. Gelation is triggered in situ by instantaneous release of ionic calci...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.21164

    authors: Reis CP,Ribeiro AJ,Neufeld RJ,Veiga F

    更新日期:2007-04-01 00:00:00

  • Promoter methylation and transgene copy numbers predict unstable protein production in recombinant Chinese hamster ovary cell lines.

    abstract::Mammalian cell lines for recombinant protein production need to maintain productivity over extended cultivation times. Long-term stability studies are time and resource intensive, but are widely performed to identify and eliminate unstable candidates during cell line development. Production instability of manufacturin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.23216

    authors: Osterlehner A,Simmeth S,Göpfert U

    更新日期:2011-11-01 00:00:00

  • The design of controllers for batch bioreactors.

    abstract::The implementation of control algorithms to batch bioreactors is often complicated by variations in process dynamics that occur during the course of fermentation. Such a wide operating range often renders the performance of fixed gain proportional-integral-differential (PID) controllers unsatisfactory. In this work, d...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260320415

    authors: Cardello RJ,San KY

    更新日期:1988-08-05 00:00:00

  • Aerobic phosphorus release linked to acetate uptake in bio-P sludge: process modeling using oxygen uptake rate.

    abstract::The main processes involved in enhanced biological phosphorus removal (EBPR) under anaerobic and subsequently aerobic conditions are widely described in the literature. Polyphosphate accumulating organisms (PAO) are the organisms responsible for this process. However, the mechanisms of PAO are not fully established ye...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10868

    authors: Guisasola A,Pijuan M,Baeza JA,Carrera J,Casas C,Lafuente J

    更新日期:2004-03-30 00:00:00

  • Large-scale expansion of human skin-derived precursor cells (hSKPs) in stirred suspension bioreactors.

    abstract::Human skin-derived precursor cells (hSKPs) are multipotent adult stem cells found in the dermis of human skin. Incorporation of hSKPs into split-thickness skin grafts (STSGs), the current gold standard to treat severe burns or tissue resections, has been proposed as a treatment option to enhance skin wound healing and...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26040

    authors: Surrao DC,Boon K,Borys B,Sinha S,Kumar R,Biernaskie J,Kallos MS

    更新日期:2016-12-01 00:00:00

  • Development of a new reversed micelle liquid emulsion membrane for protein extraction.

    abstract::A new type of liquid emulsion membrane containing reversed micelles for protein extraction is introduced. A three-step extraction mechanism is proposed including solubilization, transportation, and release of the protein. The surfactants Span80 and sodium di(2-ethylhexyl)sulfosuccinate (AOT) are used to stabilize the ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(SICI)1097-0290(19970205)53:3<267::AID-BIT

    authors: Stobbe H,Yunguang X,Zihao W,Jufu F

    更新日期:1997-02-05 00:00:00

  • Optimization of the activity in porous media of proton-generating immobilized enzymatic reactions by weak acid facilitation.

    abstract::In addition to the role of maintaining the pH, buffers can also facilitate the transport of H(+) ions in acid-generating systems. The role of this facilitation in proton transport in porous pellets on acid-generating immobilized enzymic reactions is examined. The activity in these systems can be maximized by a proper ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260270609

    authors: Ruckenstein E,Rajora P

    更新日期:1985-06-01 00:00:00

  • Maintaining data integrity in microarray data management.

    abstract::Gene expression microarrays are a relatively new technology, dating back just a few years, yet they have already become a very widely used tool in biology, and have evolved to a wide range of applications well beyond their original design intent. However, while the use of microarrays has expanded, and the issues of pe...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章,评审

    doi:10.1002/bit.10847

    authors: Grant GR,Manduchi E,Pizarro A,Stoeckert CJ Jr

    更新日期:2003-12-30 00:00:00

  • Fuzzy supervisory control of glutamic acid production.

    abstract::In glutamic acid fermentation, the molasses feeding policy and time of penicillin addition significantly affected glutamic acid production, and a fuzzy supervisory control system was developed for their quasi-optimal regulation.From the trend of the experimental data, production rules and membership functions of fuzzy...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260440113

    authors: Kitsuta Y,Kishimoto M

    更新日期:1994-06-05 00:00:00

  • Fluorescence-based sensing system for copper using genetically engineered living yeast cells.

    abstract::A whole cell-based optical sensing system for copper was developed based on Saccharomyces cerevisiae cells harboring plasmid pYEX-GFPuv. The basis of this system was the ability of the transcriptional activator protein Ace1 present in S. cerevisiae to control the expression of the reporter protein, GFPuv. When copper ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20331

    authors: Shetty RS,Deo SK,Liu Y,Daunert S

    更新日期:2004-12-05 00:00:00

  • Surface transformation of bioactive glass in bioreactors simulating microgravity conditions. Part I: experimental study.

    abstract::Surface modified bioactive glass with surface properties akin to those of the bone mineral phase is an attractive candidate for use as a microcarrier material for 3-D growth of bone-like tissue in rotating wall vessel bioreactors (RWVs). The critical surface properties of this material are the result of reaction in so...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10023

    authors: Radin S,Ducheyne P,Ayyaswamy PS,Gao H

    更新日期:2001-11-05 00:00:00

  • Dual-chambered membrane bioreactor for coculture of stratified cell populations.

    abstract::We have developed a dual-chambered bioreactor (DCB) that incorporates a membrane to study stratified 3D cell populations for skin tissue engineering. The DCB provides adjacent flow lines within a common chamber; the inclusion of the membrane regulates flow layering or mixing, which can be exploited to produce layers o...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.27164

    authors: Navarro J,Swayambunathan J,Janes ME,Santoro M,Mikos AG,Fisher JP

    更新日期:2019-12-01 00:00:00

  • Heat and mass transfer effects in static solid-substrate fermentations: design of fermentation chambers.

    abstract::An experimental device was constructed to allow nearly simultaneous measurements to be made on temperature and gas composition at different depths in a solid-substrate fermentation bed. The time-dependent values of temperature, mol % O(2) and mol % CO(2) were measured at five positions in beds 6.35 cm (2.5 in.) deep. ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260250405

    authors: Rathbun BL,Shuler ML

    更新日期:1983-04-01 00:00:00

  • Kinetic analysis of bioconversion of cellulose in attrition bioreactor.

    abstract::The attrition bioreactor (ABR) combines wet ball milling and enzymatic hydrolysis in one process step. It was found that the ABR did not accelerate enzyme deactivation. Interfacial forces, not shear forces, caused the most deactivation. Elimination of the air-liquid interface by covering the reactor substantially incr...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260310106

    authors: Jones EO,Lee JM

    更新日期:1988-01-01 00:00:00

  • Comparative studies on enzyme preparations and role of cell components for (R)-phenylacetylcarbinol production in a two-phase biotransformation.

    abstract::Whole cell pyruvate decarboxylase (PDC) from Candida utilis enhanced the enzymatic production of (R)-phenylacetylcarbinol (PAC) in an aqueous/octanol biotransformation compared to the partially purified PDC especially for a lower range of initial activities (0.3-2.5 U/mL). With an initial activity of 1.1 U/mL and at a...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20959

    authors: Satianegara G,Rogers PL,Rosche B

    更新日期:2006-08-20 00:00:00