Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells.

Abstract:

:For most cultivated mammalian cells, glutamine is an essential medium component. However, glutamine consumption results in the production of ammonia, a cytotoxic byproduct. Here we investigated the effect of glutamine reduction on recombinant protein production and ammonia accumulation in transiently transfected CHO and HEK-293E cells maintained under conditions of growth arrest. Maximum transient recombinant protein yields were observed in HEK-293E cultures without glutamine and in CHO cultures with 2 mM glutamine. The initial concentration of glutamine correlated with the level of ammonia accumulation in each culture. For both a stable CHO-derived cell line and a polyclonal population of recombinant CHO cells grown under conditions of mild hypothermia, the highest volumetric protein productivity was observed in cultures without glutamine. Here, the level of ammonia accumulation also corresponded to the initial glutamine concentration. Our data demonstrate that reduction of glutamine in the medium is an effective approach to improve protein production in both transiently and stably transfected mammalian cells when applying conditions that reduce or arrest the growth of these cells.

journal_name

Biotechnol Lett

journal_title

Biotechnology letters

authors

Rajendra Y,Kiseljak D,Baldi L,Hacker DL,Wurm FM

doi

10.1007/s10529-011-0809-z

subject

Has Abstract

pub_date

2012-04-01 00:00:00

pages

619-26

issue

4

eissn

0141-5492

issn

1573-6776

journal_volume

34

pub_type

杂志文章
  • Evaluation of inert and organic carriers for Verticillium lecanii spore production in solid-state fermentation.

    abstract::Growth and sporulation of Verticillium lecanii on inert and organic carriers (sugar-cane bagasse, corncob, rice straw, polyurethane foam and activated carbon) in a solid-state fermentation process was studied. Sugar-cane bagasse and polyurethane foam produced 10(10) spores g(-1) dry carrier whereas corncob, rice straw...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-010-0496-1

    authors: Xu X,Yu Y,Shi Y

    更新日期:2011-04-01 00:00:00

  • High-level expression of functional recombinant human coagulation factor VII in insect cells.

    abstract::Recombinant coagulation factor VII (FVII) is used as a potential therapeutic intervention in hemophilia patients who produce antibodies against the coagulation factors. Mammalian cell lines provide low levels of expression, however, the Spodoptera frugiperda Sf9 cell line and baculovirus expression system are powerful...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-010-0227-7

    authors: Masroori N,Halabian R,Mohammadipour M,Roushandeh AM,Rouhbakhsh M,Najafabadi AJ,Fathabad ME,Salimi M,Shokrgozar MA,Roudkenar MH

    更新日期:2010-06-01 00:00:00

  • 4-Vinylphenol production from glucose using recombinant Streptomyces mobaraense expressing a tyrosine ammonia lyase from Rhodobacter sphaeroides.

    abstract:OBJECTIVES:To find a novel host for the production of 4-vinylphenol (4VPh) by screening Streptomyces species. RESULTS:The conversion of p-coumaric acid (pHCA) to 4VPh in Streptomyces mobaraense was evaluated using a medium containing pHCA. S. mobaraense readily assimilated pHCA after 24 h of cultivation to produce 4VP...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-016-2126-z

    authors: Fujiwara R,Noda S,Kawai Y,Tanaka T,Kondo A

    更新日期:2016-09-01 00:00:00

  • Cloning, expression and characterization of glycoside hydrolase family 11 endoxylanase from Bacillus pumilus ARA.

    abstract::An endoxylanase gene, xynA, was cloned from Bacillus pumilus ARA and expressed in Escherichia coli. The open reading frame of the xynA gene was 687 bp encoding a signal peptide and a mature xylanase with a molecular mass of 23 kDa. The enzyme was categorized as a glycosyl hydrolase family 11 member based on the sequen...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-011-0568-x

    authors: Qu W,Shao W

    更新日期:2011-07-01 00:00:00

  • Design of acoustic cell settler for filtering and recycling microbial cells.

    abstract::An acoustic cell settler (ACS) using ultrasound at cells of 3 MHz was used to recycle Saccharomyces cerevisiae in a fermenter. The locations of both the inlet and outlet in the acoustic cell settler, which have a relatively long distance between the transducer and reflector, were optimized. A tilted settler was design...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1023/a:1022345508979

    authors: Hwang SH,Koo YM

    更新日期:2003-02-01 00:00:00

  • Heterologous expression of human interleukin-6 in Streptomyces lividans TK24 using novel secretory expression vectors.

    abstract::Streptomyces is an attractive host for heterologous protein secretion. To further optimize its expression capacity, better expression vectors will be helpful. Here, based on pSGL1, a high copy number plasmid present in Streptomyces globisporus C-1027, we constructed a series of novel E. coli-Streptomyces shuttle expre...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-010-0428-0

    authors: Zhu Y,Wang L,Du Y,Wang S,Yu T,Hong B

    更新日期:2011-02-01 00:00:00

  • Induced hydrolytic enzymes of ectomycorrhizal fungi against pathogen Rhizoctonia solani.

    abstract::Interactions between ectomycorrhizal fungi (Suillus laricinus, S. tomentosus, Amanita vaginata and Gomphidius viscidus) and the pathogen Rhizoctonia solani in co-culture were studied using both light and scanning electron microscopy. S. laricinus, S. tomentosus and A. vaginata inhibited the growth of the pathogen. Mor...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-008-9760-z

    authors: Tang M,Zhang RQ,Chen H,Zhang HH,Tian ZQ

    更新日期:2008-10-01 00:00:00

  • Generation of ΔF508-CFTR T84 cell lines by CRISPR/Cas9-mediated genome editing.

    abstract:OBJECTIVES:To provide a simple method to make a stable ΔF508-CFTR-expressing T84 cell line that can be used as an efficient screening model system for ΔF508-CFTR rescue. RESULTS:CFTR knockout cell lines were generated by Cas9 with a single-guide RNA (sgRNA) targeting exon 1 of the CFTR genome, which produced indels th...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-016-2190-4

    authors: Chung WY,Song M,Park J,Namkung W,Lee J,Kim H,Lee MG,Kim JY

    更新日期:2016-12-01 00:00:00

  • Unexpected reaction profile observed in the synthesis of propyl laurate when using Candida rugosa lipases immobilized in microemulsions based organogels.

    abstract::1-Propyl laurate synthesis should not be used as standard reaction test of immobilized enzymes in microemulsion-based organogels (MBGs) prepared using lecithin/1-propanol as surfactant when extremely active enzymes with high load are used. In these cases, an anomalous kinetic reaction constant value is observed over s...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1023/B:BILE.0000044455.42471.bf

    authors: Domínguez de María P,Xenakis A,Stamatis H,Sinisterra JV

    更新日期:2004-10-01 00:00:00

  • Stabilization of human immunodeficiency virus type 1 reverse transcriptase by site-directed mutagenesis.

    abstract::Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is a heterodimer containing 66 kDa p66 and 51 kDa p51 subunits. We previously showed that HIV-1 group M (HIV-1 M) RT and HIV-1 group O (HIV-1 O) RT have higher affinities for dTTP and template-primer (T/P) than Moloney murine leukemia virus RT, whi...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-013-1321-4

    authors: Nishimura K,Shinomura M,Konishi A,Yasukawa K

    更新日期:2013-12-01 00:00:00

  • Bioreactors for tissue engineering.

    abstract::Bioreactors are essential in tissue engineering, not only because they provide an in vitro environment mimicking in vivo conditions for the growth of tissue substitutes, but also because they enable systematic studies of the responses of living tissues to various mechanical and biochemical cues. The basic principles o...

    journal_title:Biotechnology letters

    pub_type: 杂志文章,评审

    doi:10.1007/s10529-006-9111-x

    authors: Chen HC,Hu YC

    更新日期:2006-09-01 00:00:00

  • Potential of biodegradable microneedles as a transdermal delivery vehicle for lidocaine.

    abstract::There has been an increasing interest in applying biotechnology in formulating and characterising new and innovative drug delivery methods, e.g., drug-loaded biodegradable microneedles within the area of transdermal delivery technology. Recently, microneedles have been proposed for use in pain management, e.g., post-o...

    journal_title:Biotechnology letters

    pub_type: 杂志文章,评审

    doi:10.1007/s10529-013-1217-3

    authors: Nayak A,Das DB

    更新日期:2013-09-01 00:00:00

  • (-)-Epicatechin-3-gallate (a polyphenol from green tea) potentiates doxorubicin-induced apoptosis in H9C2 cardiomyocytes.

    abstract:OBJECTIVE:The effect of epicatechin-3-gallate (ECG), a polyphenol that is present in green tea, on doxorubicin (DOX) cytotoxicity in H9C2 cardiomyocytes and its underlying mechanisms were investigated. RESULTS:Pretreatment with ECG (20 and 30 μM) significantly increased DOX-induced apoptosis to 16-18% in H9C2 cardiomy...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-015-1879-0

    authors: Cai Y,Hong HQ,Zhao L,Cai YP,Qin Y

    更新日期:2015-10-01 00:00:00

  • A rapid and efficient polyethylenimine-based transfection method to prepare lentiviral or retroviral vectors: useful for making iPS cells and transduction of primary cells.

    abstract:OBJECTIVES:To improve the efficiency, reproducibility and consistency of the PEI-based transfection method that is often used in preparation of recombinant lentiviral or retroviral vectors. RESULTS:The contributions to transfection efficiency of multi-factors including concentration of PEI or DNA, dilution buffer for ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-016-2123-2

    authors: Yang S,Shi H,Chu X,Zhou X,Sun P

    更新日期:2016-09-01 00:00:00

  • Alginate oligosaccharide protects endothelial cells against oxidative stress injury via integrin-α/FAK/PI3K signaling.

    abstract::Alginate oligosaccharide (AOS) was reported to possess antioxidant and free radical scavenging activities, but the specific effects and mechanisms remain unclear. We investigated the effects of AOS on H2O2-induced oxidative stress and apoptosis in human umbilical vein endothelial cells (HUVECs) and the associated mech...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-020-03010-z

    authors: Zhao J,Han Y,Wang Z,Zhang R,Wang G,Mao Y

    更新日期:2020-12-01 00:00:00

  • Kinetic mechanism of streptomycin adenylyltransferase from a recombinant Escherichia coli.

    abstract::Bacterial resistance to the aminoglycoside antibiotics is manifested primarily by enzymic modification of these drugs. One important mechanism of streptomycin modification is through ATP-dependent O-adenylation, catalyzed by streptomycin adenylyltransferase. Initial velocity patterns deduced from steady state kinetics...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-005-2544-9

    authors: Jana S,Deb JK

    更新日期:2005-04-01 00:00:00

  • Design of a multi-enzyme reaction on an electrode surface for an L-glutamate biofuel anode.

    abstract:OBJECTIVES:To design and construct a novel bio-anode electrode based on the oxidation of glutamic acid to produce 2-oxoglutarate, generating two electrons from NADH. RESULTS:Efficient enzyme reaction and electron transfer were observed owing to immobilization of the two enzymes using a mixed self-assembled monolayer. ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-016-2237-6

    authors: Sakamoto H,Komatsu T,Yamasaki K,Satomura T,Suye SI

    更新日期:2017-02-01 00:00:00

  • Increased expression of OsPT1, a high-affinity phosphate transporter, enhances phosphate acquisition in rice.

    abstract::Most high-affinity phosphate transporter genes (OsPTs) in rice were highly induced in roots when phosphate was depleted. OsPT1, however, was highly expressed in primary roots and leaves regardless of external phosphate concentrations. This finding was confirmed histochemically using transgenic rice plants that express...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-008-9757-7

    authors: Seo HM,Jung Y,Song S,Kim Y,Kwon T,Kim DH,Jeung SJ,Yi YB,Yi G,Nam MH,Nam J

    更新日期:2008-10-01 00:00:00

  • Construction of streptavidin-luciferase fusion protein for ATP sensing with fixed form.

    abstract::A fusion protein consisting of streptavidin and firefly luciferase was constructed to establish an accurate measuring technique of local ATP concentration. The fusion protein retained the binding ability of streptavidin and enzymatic activity of luciferase. Also, it could detect the concentration of antigens and could...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1023/B:BILE.0000032966.17759.08

    authors: Nakamura M,Mie M,Funabashi H,Kobatake E

    更新日期:2004-07-01 00:00:00

  • Conserved microRNA function as a basis for Chinese hamster ovary cell engineering.

    abstract::The use of microRNAs (miRNAs) for improving the efficiency of recombinant protein production by CHO cells is gaining considerable interest for their ability to regulate entire molecular networks. Differential miRNA expression profiling and large-scale transient screening have been the prerequisite for the selection of...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-014-1751-7

    authors: Kelly PS,Gallagher C,Clynes M,Barron N

    更新日期:2015-04-01 00:00:00

  • Strategy for enhancing adenosine production under the guidance of transcriptional and metabolite pool analysis.

    abstract:OBJECTIVES:To rationally identify targets for enhancing adenosine production, transcription level of genes involved in adenosine synthesis of Bacillus subtilis XGL was detected during the fermentation process, complemented with metabolite pool analysis. RESULTS:PurR-regulated genes (pur operon and purA) and prs were d...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-015-1801-9

    authors: Zhang C,Du S,Liu Y,Xie X,Xu Q,Chen N

    更新日期:2015-07-01 00:00:00

  • Characterization of ribose-5-phosphate isomerase of Clostridium thermocellum producing D-allose from D-psicose.

    abstract::The rpiB gene, encoding ribose-5-phosphate isomerase (RpiB) from Clostridium thermocellum, was cloned and expressed in Escherichia coli. RpiB converted D-psicose into D-allose but it did not convert D-xylose, L-rhamnose, D-altrose or D-galactose. The production of D-allose by RpiB was maximal at pH 7.5 and 65 degrees ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-007-9393-7

    authors: Park CS,Yeom SJ,Kim HJ,Lee SH,Lee JK,Kim SW,Oh DK

    更新日期:2007-09-01 00:00:00

  • Anti-prelog reduction of prochiral carbonyl compounds by Oenococcus oeni in a biphasic system.

    abstract::An aqueous-organic biphasic system was established and used with whole cells of Oenococcus oeni to reduce 2-octanone to (R)-2-octanol. The conversion reached 99% when the Tris/borate buffer was increased from 50 mM to 300 mM in the aqueous phase. In addition, the conversion increased as the log P value of the organic ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-006-9062-2

    authors: Hu J,Xu Y

    更新日期:2006-07-01 00:00:00

  • Improved cellobiose utilization in E. coli by including both hydrolysis and phosphorolysis mechanisms.

    abstract::Cellobiose is a major intermediate from cellulase hydrolysis of pretreated plant biomass. Engineering biocatalysts for direct use of cellobiose could eliminate the need for exogenous β-glucosidase. Additionally, rapid removal of cellobiose in a simultaneous saccharification and fermentation facilitates enzymatic hydro...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-013-1355-7

    authors: Rutter C,Chen R

    更新日期:2014-02-01 00:00:00

  • Expression of the human cancer/testis antigen NY-SAR-35 is activated by CpG island hypomethylation.

    abstract::The novel cancer/testis antigen gene, NY-SAR-35, is expressed exclusively in normal testis and in various histological types of tumor. However, the NY-SAR-35 gene expression is observed to be aberrant in several cancer cell lines and tissues. The analysis of methylation status of the NY-SAR-35 gene promoter in various...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-011-0559-y

    authors: Park JH,Song MH,Lee CH,Lee MK,Park YM,Old L,Lee SY

    更新日期:2011-06-01 00:00:00

  • A polysaccharide isolated from the medicinal herb Bletilla striata induces endothelial cells proliferation and vascular endothelial growth factor expression in vitro.

    abstract::A polysaccharide from the traditional Chinese medicinal herb, Bletilla striata (Thunb.) Reichb. f., was isolated, purified and characterized. It induced the proliferation of human umbilical vascular endothelial cells and the expression of vascular endothelial growth factor up to 156% and 147% of control after 72 h, re...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-006-0011-x

    authors: Wang C,Sun J,Luo Y,Xue W,Diao H,Dong L,Chen J,Zhang J

    更新日期:2006-04-01 00:00:00

  • A novel two-step fermentation process for improved arachidonic acid production by Mortierella alpina.

    abstract::A novel two-step fermentation process was developed to enhance arachidonic acid (ARA) production by Mortierella alpina ME-1 in a 5 l fermentor. Agitation speed and aeration rate were adjusted from 180 to 40 rpm and from 0.6 to 1 vvm, respectively, after 5 days cultivation, to decrease physical damage to the mycelia an...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-008-9661-1

    authors: Jin MJ,Huang H,Xiao AH,Zhang K,Liu X,Li S,Peng C

    更新日期:2008-06-01 00:00:00

  • Involvement of polyamines in plant response to abiotic stress.

    abstract::Environmental stresses are the major cause of crop loss worldwide. Polyamines are involved in plant stress responses. However, the precise role(s) of polyamine metabolism in these processes remain ill-defined. Transgenic approaches demonstrate that polyamines play essential roles in stress tolerance and open up the po...

    journal_title:Biotechnology letters

    pub_type: 杂志文章,评审

    doi:10.1007/s10529-006-9179-3

    authors: Alcázar R,Marco F,Cuevas JC,Patron M,Ferrando A,Carrasco P,Tiburcio AF,Altabella T

    更新日期:2006-12-01 00:00:00

  • Deletion of a KU80 homolog enhances homologous recombination in the thermotolerant yeast Kluyveromyces marxianus.

    abstract::Targeted gene replacement in the thermotolerant yeast Kluyveromyces marxianus KCTC 17555 has been hampered by its propensity to non-homologous end joining (NHEJ). To enhance homologous recombination (HR) by blocking NHEJ, we identified and disrupted the K. marxianus KU80 gene. The ku80 deletion mutant strain (Kmku80∆)...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-014-1576-4

    authors: Choo JH,Han C,Kim JY,Kang HA

    更新日期:2014-10-01 00:00:00

  • Constructing an improved pyrroloquinoline quinone glucose dehydrogenase binding aptamer for enzyme labeling.

    abstract::Motif analysis of PGa4 indicated that eight consecutive guanine residues were most important for pyrroloquinoline quinine glucose dehydrogenase (PQQGDH) recognition. Because the truncated mutants minimized unexpected interactions between PGa4 and the target-binding aptamer, they will now facilitate the construction of...

    journal_title:Biotechnology letters

    pub_type: 杂志文章

    doi:10.1007/s10529-010-0292-y

    authors: Abe K,Sode K,Ikebukuro K

    更新日期:2010-09-01 00:00:00