Penicillin production in industrial strain Penicillium chrysogenum P2niaD18 is not dependent on the copy number of biosynthesis genes.

Abstract:

BACKGROUND:Multi-copy gene integration into microbial genomes is a conventional tool for obtaining improved gene expression. For Penicillium chrysogenum, the fungal producer of the beta-lactam antibiotic penicillin, many production strains carry multiple copies of the penicillin biosynthesis gene cluster. This discovery led to the generally accepted view that high penicillin titers are the result of multiple copies of penicillin genes. Here we investigated strain P2niaD18, a production line that carries only two copies of the penicillin gene cluster. RESULTS:We performed pulsed-field gel electrophoresis (PFGE), quantitative qRT-PCR, and penicillin bioassays to investigate production, deletion and overexpression strains generated in the P. chrysogenum P2niaD18 background, in order to determine the copy number of the penicillin biosynthesis gene cluster, and study the expression of one penicillin biosynthesis gene, and the penicillin titer. Analysis of production and recombinant strain showed that the enhanced penicillin titer did not depend on the copy number of the penicillin gene cluster. Our assumption was strengthened by results with a penicillin null strain lacking pcbC encoding isopenicillin N synthase. Reintroduction of one or two copies of the cluster into the pcbC deletion strain restored transcriptional high expression of the pcbC gene, but recombinant strains showed no significantly different penicillin titer compared to parental strains. CONCLUSIONS:Here we present a molecular genetic analysis of production and recombinant strains in the P2niaD18 background carrying different copy numbers of the penicillin biosynthesis gene cluster. Our analysis shows that the enhanced penicillin titer does not strictly depend on the copy number of the cluster. Based on these overall findings, we hypothesize that instead, complex regulatory mechanisms are prominently implicated in increased penicillin biosynthesis in production strains.

journal_name

BMC Biotechnol

journal_title

BMC biotechnology

authors

Ziemons S,Koutsantas K,Becker K,Dahlmann T,Kück U

doi

10.1186/s12896-017-0335-8

subject

Has Abstract

pub_date

2017-02-16 00:00:00

pages

16

issue

1

issn

1472-6750

pii

10.1186/s12896-017-0335-8

journal_volume

17

pub_type

杂志文章
  • Production of recombinant VP1-derived virus-like particles from novel human polyomaviruses in yeast.

    abstract:BACKGROUND:Eleven new human polyomaviruses (HPyVs) have been identified in the last decade. Serological studies show that these novel HPyVs sub-clinically infect humans at an early age. The routes of infection, entry pathways, and cell tropism of new HPyVs remain unknown. VP1 proteins of polyomaviruses can assembly int...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0187-z

    authors: Norkiene M,Stonyte J,Ziogiene D,Mazeike E,Sasnauskas K,Gedvilaite A

    更新日期:2015-08-04 00:00:00

  • The relationship between mTOR signalling pathway and recombinant antibody productivity in CHO cell lines.

    abstract:BACKGROUND:High recombinant protein productivity in mammalian cell lines is often associated with phenotypic changes in protein content, energy metabolism, and cell growth, but the key determinants that regulate productivity are still not clearly understood. The mammalian target of rapamycin (mTOR) signalling pathway h...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-14-15

    authors: Edros R,McDonnell S,Al-Rubeai M

    更新日期:2014-02-17 00:00:00

  • Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes.

    abstract:BACKGROUND:The CRISPR/Cas9 system is increasingly used for gene inactivation in mouse zygotes, but homology-directed mutagenesis and use of inbred embryos are less established. In particular, Rosa26 knock-in alleles for the insertion of transgenes in a genomic 'safe harbor' site, have not been produced. Here we applied...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-016-0234-4

    authors: Chu VT,Weber T,Graf R,Sommermann T,Petsch K,Sack U,Volchkov P,Rajewsky K,Kühn R

    更新日期:2016-01-16 00:00:00

  • Pattern analysis approach reveals restriction enzyme cutting abnormalities and other cDNA library construction artifacts using raw EST data.

    abstract:BACKGROUND:Expressed Sequence Tag (EST) sequences are widely used in applications such as genome annotation, gene discovery and gene expression studies. However, some of GenBank dbEST sequences have proven to be "unclean". Identification of cDNA termini/ends and their structures in raw ESTs not only facilitates data qu...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-12-16

    authors: Zhou S,Ji G,Liu X,Li P,Moler J,Karro JE,Liang C

    更新日期:2012-05-03 00:00:00

  • Effect of the cancer specific shorter form of human 6-phosphofructo-1-kinase on the metabolism of the yeast Saccharomyces cerevisiae.

    abstract:BACKGROUND:At first glance, there appears to be a high degree of similarity between the metabolism of yeast (the Crabtree effect) and human cancer cells (the Warburg effect). At the root of both effects is accelerated metabolic flow through glycolysis which leads to overflows of ethanol and lactic acid, respectively. I...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-017-0362-5

    authors: Andrejc D,Možir A,Legiša M

    更新日期:2017-05-08 00:00:00

  • A new single-step protocol for rapid baculovirus-driven protein production in insect cells.

    abstract:BACKGROUND:In the last three decades, the Baculovirus expression vector system (BEV) has evolved to one of the most widely used eukaryotic systems for heterologous protein expression including approved vaccines and therapies. Despite the significant improvements introduced during the past years, the BEV system still ha...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-017-0400-3

    authors: Scholz J,Suppmann S

    更新日期:2017-11-16 00:00:00

  • Selection of oleaginous yeasts for fatty acid production.

    abstract:BACKGROUND:Oleaginous yeast species are an alternative for the production of lipids or triacylglycerides (TAGs). These yeasts are usually non-pathogenic and able to store TAGs ranging from 20 % to 70 % of their cell mass depending on culture conditions. TAGs originating from oleaginous yeasts can be used as the so-call...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-016-0276-7

    authors: Lamers D,van Biezen N,Martens D,Peters L,van de Zilver E,Jacobs-van Dreumel N,Wijffels RH,Lokman C

    更新日期:2016-05-27 00:00:00

  • Improvement of an antibody-enzyme coupling yield by enzyme surface supercharging.

    abstract:BACKGROUND:Protein cross-coupling reactions demand high yields, especially if the products are intended for bioanalytics, like enzyme-linked immunosorbent assays. Amongst other factors, the coupling yield depends on the concentration of the proteins being used for coupling. Protein supercharging of enzymes can increase...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-014-0088-6

    authors: Prasse AA,Zauner T,Büttner K,Hoffmann R,Zuchner T

    更新日期:2014-10-18 00:00:00

  • Genetic transformation of einkorn (Triticum monococcum L. ssp. monococcum L.), a diploid cultivated wheat species.

    abstract:BACKGROUND:Domesticated einkorn (Triticum monococcum L.) is one of the oldest cultivated cereal crops in the world. Its small genome size (~ 5.7 GB), low ploidy (2n = 2x = 14, AmAm) and high genetic polymorphism make this species very attractive for use as a diploid model for understanding the genomics and proteomics o...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-018-0477-3

    authors: Miroshnichenko D,Ashin D,Pushin A,Dolgov S

    更新日期:2018-10-23 00:00:00

  • Secretion of Fc-amidated peptide fusion proteins by Chinese hamster ovary cells.

    abstract:BACKGROUND:The therapeutic use of α-amidated peptides (e.g. calcitonin, glucagon-like peptide) has increased dramatically, but there are major impediments to wider use of such peptides. Larger peptides are expensive to synthesize, and short plasma half-lives frequently limit the clinical circumstances in which the pept...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0173-5

    authors: Carlson KR,Pomerantz SC,Li J,Vafa O,Naso M,Strohl W,Mains RE,Eipper BA

    更新日期:2015-06-27 00:00:00

  • Liposome retention in size exclusion chromatography.

    abstract:BACKGROUND:Size exclusion chromatography is the method of choice for separating free from liposome-encapsulated molecules. However, if the column is not presaturated with lipids this type of chromatography causes a significant loss of lipid material. To date, the mechanism of lipid retention is poorly understood. It ha...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-5-11

    authors: Ruysschaert T,Marque A,Duteyrat JL,Lesieur S,Winterhalter M,Fournier D

    更新日期:2005-05-10 00:00:00

  • Multi-substrate chromosome preparations for high throughput comparative FISH.

    abstract:BACKGROUND:A modification of a standard method of fluorescence in situ hybridisation (FISH) is described, by which a combination of several substrates and probes on single microscope slides enables more accurate comparisons of the distribution and abundance of chromosomal sequences and improves the relatively low throu...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-6-20

    authors: Hasterok R,Dulawa J,Jenkins G,Leggett M,Langdon T

    更新日期:2006-03-20 00:00:00

  • A novel psychrophilic alkaline phosphatase from the metagenome of tidal flat sediments.

    abstract:BACKGROUND:Alkaline phosphatase (AP) catalyzes the hydrolytic cleavage of phosphate monoesters under alkaline conditions and plays important roles in microbial ecology and molecular biology applications. Here, we report on the first isolation and biochemical characterization of a thermolabile AP from a metagenome. RES...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0115-2

    authors: Lee DH,Choi SL,Rha E,Kim SJ,Yeom SJ,Moon JH,Lee SG

    更新日期:2015-01-31 00:00:00

  • Novel anti-CD38 humanized mAb SG003 possessed enhanced cytotoxicity in lymphoma than Daratumumab via antibody-dependent cell-mediated cytotoxicity.

    abstract:BACKGROUND:In vivo use of monoclonal antibodies has become routine clinical practice in the treatment of human cancer. CD38 is an attractive target, because it has double roles, as a receptor and an ectoenzyme. Daratumumab, an anti-CD38 antibody, is currently in the clinical trials for multiple myeloma. RESULTS:Here w...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-019-0524-8

    authors: Yu T,Qiao C,Lv M,Tang L

    更新日期:2019-05-22 00:00:00

  • A novel carboxyl-terminal protease derived from Paenibacillus lautus CHN26 exhibiting high activities at multiple sites of substrates.

    abstract:BACKGROUND:Carboxyl-terminal protease (CtpA) plays essential functions in posttranslational protein processing in prokaryotic and eukaryotic cells. To date, only a few bacterial ctpA genes have been characterized. Here we cloned and characterized a novel CtpA. The encoding gene, ctpAp (ctpA of Paenibacillus lautus), wa...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-13-89

    authors: Li Y,Pan Y,She Q,Chen L

    更新日期:2013-10-25 00:00:00

  • Optimization of a nicotine degrading enzyme for potential use in treatment of nicotine addiction.

    abstract:BACKGROUND:Smoking and tobacco use continue to be the largest preventable causes of death globally. A novel therapeutic approach has recently been proposed: administration of an enzyme that degrades nicotine, the main addictive component of tobacco, minimizing brain exposure and reducing its reinforcing effects. Pre-cl...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-019-0551-5

    authors: Thisted T,Biesova Z,Walmacq C,Stone E,Rodnick-Smith M,Ahmed SS,Horrigan SK,Van Engelen B,Reed C,Kalnik MW

    更新日期:2019-08-02 00:00:00

  • Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes.

    abstract:BACKGROUND:Xenoantigens are a major source of concern with regard to the success of interspecific xenografts. GGTA1 encodes α1,3-galactosyltransferase, which is essential for the biosynthesis of galactosyl-alpha 1,3-galactose, the major xenoantigen causing hyperacute rejection. GGTA1-modified pigs, therefore, are promi...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-020-00638-7

    authors: Tanihara F,Hirata M,Nguyen NT,Sawamoto O,Kikuchi T,Doi M,Otoi T

    更新日期:2020-08-18 00:00:00

  • Stable high-level expression of factor VIII in Chinese hamster ovary cells in improved elongation factor-1 alpha-based system.

    abstract:BACKGROUND:Recombinant factor VIII (FVIII), used for haemophilia A therapy, is one of the most challenging among the therapeutic proteins produced in heterologous expression systems. Deletion variant of FVIII, in which the entire domain B is replaced by a short linker peptide, was approved for medical use. Efficacy and...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-017-0353-6

    authors: Orlova NA,Kovnir SV,Gabibov AG,Vorobiev II

    更新日期:2017-03-24 00:00:00

  • The case for well-conducted experiments to validate statistical protocols for 2D gels: different pre-processing = different lists of significant proteins.

    abstract:BACKGROUND:The proteomics literature has seen a proliferation of publications that seek to apply the rapidly improving technology of 2D gels to study various biological systems. However, there is a dearth of systematic studies that have investigated appropriate statistical approaches to analyse the data from these expe...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-5-7

    authors: Meleth S,Deshane J,Kim H

    更新日期:2005-02-11 00:00:00

  • Tetanus toxin fragments and Bcl-2 fusion proteins: cytoprotection and retrograde axonal migration.

    abstract:BACKGROUND:Tetanus neurotoxin (TeNT) is taken up at nerve terminals and undergoes retrograde migration. The toxic properties of TeNT reside in the toxin light chain (L), but like complete TeNT, the TeNT heavy chain (TTH) and the C-terminal domain (TTC) alone can bind and enter into neurons. Here, we explored whether at...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-018-0452-z

    authors: Watanabe Y,Matsuba T,Nakanishi M,Une M,Hanajima R,Nakashima K

    更新日期:2018-06-11 00:00:00

  • Description of a PCR-based technique for DNA splicing and mutagenesis by producing 5' overhangs with run through stop DNA synthesis utilizing Ara-C.

    abstract:BACKGROUND:Splicing of DNA molecules is an important task in molecular biology that facilitates cloning, mutagenesis and creation of chimeric genes. Mutagenesis and DNA splicing techniques exist, some requiring restriction enzymes, and others utilize staggered reannealing approaches. RESULTS:A method for DNA splicing ...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-5-23

    authors: Ailenberg M,Goldenberg NM,Silverman M

    更新日期:2005-09-01 00:00:00

  • Recombinant mussel proximal thread matrix protein promotes osteoblast cell adhesion and proliferation.

    abstract:BACKGROUND:von Willebrand factor (VWF) is a key load bearing domain for mamalian cell adhesion by binding various macromolecular ligands in extracellular matrix such as, collagens, elastin, and glycosaminoglycans. Interestingly, vWF like domains are also commonly found in load bearing systems of marine organisms such a...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-016-0247-z

    authors: Yoo HY,Song YH,Foo M,Seo E,Hwang DS,Seo JH

    更新日期:2016-02-16 00:00:00

  • Systematic cross-validation of 454 sequencing and pyrosequencing for the exact quantification of DNA methylation patterns with single CpG resolution.

    abstract:BACKGROUND:New high-throughput sequencing technologies promise a very sensitive and high-resolution analysis of DNA methylation patterns in quantitative terms. However, a detailed and comprehensive comparison with existing validated DNA methylation analysis methods is not yet available. Therefore, a systematic cross-va...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-11-6

    authors: Potapova A,Albat C,Hasemeier B,Haeussler K,Lamprecht S,Suerbaum S,Kreipe H,Lehmann U

    更新日期:2011-01-14 00:00:00

  • A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions.

    abstract:BACKGROUND:Analysis of protein-protein interactions (PPIs) is a valuable approach for the characterization of huge networks of protein complexes or proteins of unknown function. Co-immunoprecipitation (coIP) using affinity resins coupled to protein A/G is the most widely used method for PPI detection. However, this tra...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-10-78

    authors: Liu Q,Chen Q,Wang J,Zhang Y,Zhou Y,Lin C,He W,He F,Xu D

    更新日期:2010-10-28 00:00:00

  • Expression of Vitreoscilla hemoglobin enhances production of arachidonic acid and lipids in Mortierella alpina.

    abstract:BACKGROUND:Arachidonic acid (ARA, C20:4, n-6), which belongs to the omega-6 series of polyunsaturated fatty acids and has a variety of biological activities, is commercially produced in Mortierella alpina. Dissolved oxygen or oxygen utilization efficiency is a critical factor for Mortierella alpina growth and arachidon...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-017-0388-8

    authors: Zhang H,Feng Y,Cui Q,Song X

    更新日期:2017-08-30 00:00:00

  • Pharmacological characteristics and efficacy of a novel anti-angiogenic antibody FD006 in corneal neovascularization.

    abstract:BACKGROUND:Vascular endothelial growth factor (VEGF) is a key angiogenic factors. It plays an important role in both physiologic and pathologic angiogenesis and increases permeability across the vessels. Using antibody phage display technology, we obtained a novel anti-VEGFA IgG, named as FD006. In this study, the phar...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-14-17

    authors: Wang Q,Yang J,Tang K,Luo L,Wang L,Tian L,Jiang Y,Feng J,Li Y,Shen B,Lv M,Huang Y

    更新日期:2014-02-27 00:00:00

  • CRISPR/Cas9-mediated genome editing induces gene knockdown by altering the pre-mRNA splicing in mice.

    abstract:BACKGROUND:Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) has been wildly used to generate gene knockout models through inducing indels causing frame-shift. However, there are few studies concerning the post-transcript effects caused by CRISPR-mediated genome editing...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-018-0472-8

    authors: Tang JX,Chen D,Deng SL,Li J,Li Y,Fu Z,Wang XX,Zhang Y,Chen SR,Liu YX

    更新日期:2018-10-03 00:00:00

  • Selective permeation and organic extraction of recombinant green fluorescent protein (gfpuv) from Escherichia coli.

    abstract:BACKGROUND:Transformed cells of Escherichia coli DH5-alpha with pGFPuv, induced by IPTG (isopropyl-beta-d-thiogalactopyranoside), express the green fluorescent protein (gfpuv) during growth phases. E. coli subjected to the combination of selective permeation by freezing/thawing/sonication cycles followed by the three-p...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-2-7

    authors: Vessoni Penna TC,Ishii M

    更新日期:2002-04-24 00:00:00

  • A new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) affects Soybean Asian rust (Phakopsora pachyrhizi) spore germination.

    abstract:BACKGROUND:Asian rust (Phakopsora pachyrhizi) is a common disease in Brazilian soybean fields and it is difficult to control. To identify a biochemical candidate with potential to combat this disease, a new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) (CaclXIP) leaves was cloned into the...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-11-14

    authors: Vasconcelos EA,Santana CG,Godoy CV,Seixas CD,Silva MS,Moreira LR,Oliveira-Neto OB,Price D,Fitches E,Filho EX,Mehta A,Gatehouse JA,Grossi-De-Sa MF

    更新日期:2011-02-07 00:00:00

  • In vitro cell cultures obtained from different explants of Corylus avellana produce Taxol and taxanes.

    abstract:BACKGROUND:Taxol is an effective antineoplastic agent, originally extracted from the bark of Taxus brevifolia with a low yield. Many attempts have been made to produce Taxol by chemical synthesis, semi-synthesis and plant tissue cultures. However, to date, the availability of this compound is not sufficient to satisfy ...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-6-45

    authors: Bestoso F,Ottaggio L,Armirotti A,Balbi A,Damonte G,Degan P,Mazzei M,Cavalli F,Ledda B,Miele M

    更新日期:2006-12-06 00:00:00