Cloning of the SAP6 gene of Metschnikowia reukaufii and its heterologous expression and characterization in Escherichia coli.


:The SAP6 gene (without signal sequence) encoding Metschnikowia reukaufii acid protease was amplified by PCR and fused to the expression vector pET-24a(+). The carboxy-terminal 6x His-tagged recombinant acid protease (rSAP6) was expressed from pET-24a(+)SAP6-6His in Escherichia coli BL21 (DE3) and purified with affinity chromatography using a Ni-NTA column. SDS-PAGE analysis and Western blotting revealed that the molecular mass of the purified rSAP6 was 54kDa. The optimal temperature and pH of the purified rSAP6 were 40 degrees C and 3.4, respectively. The enzyme was stable below 45 degrees C and between pH 2.6 and 5.0. The results show that Mn(2+) had an activating effect on the enzyme, while Cu(2+), Mg(2+), Zn(2+) and Ag(+) acted as inhibitors of the enzyme. However, Ca(2+) had no effect on the enzyme activity. The purified rSAP6 was characterized as an aspartic protease as it was inhibited by aspartic protease-specific inhibitors, such as pepstatin. It was also found that the purified rSAP6 had milk-clotting activity.


Microbiol Res


Microbiological research


Li J,Chi Z,Wang X




Has Abstract


2010-03-31 00:00:00














  • Fe(III) reduction activity and cytochrome content of Shewanella putrefaciens grown on ten compounds as sole terminal electron acceptor.

    abstract::Shewanella putrefaciens was grown on a series of ten alternate compounds as sole terminal electron acceptor. Each cell type was analyzed for Fe(III) reduction activity, absorbance maxima in reduced-minus-oxidized difference spectra and heme-containing protein content. High-rate Fe(III) reduction activity, pronounced d...

    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Blakeney MD,Moulaei T,DiChristina TJ

    更新日期:2000-07-01 00:00:00

  • Characterization of imipenem-resistant Serratia marcescens producing IMP-type and TEM-type beta-lactamases encoded on a single plasmid.

    abstract::To evaluate the roles of blaIMP and blaTEM genes in the resistance of Serratia marcescens against beta-lactams and to find the spreading ways of these genes, 19 clinical isolates of imipenem-resistant Serratia marcescens were analyzed. Six strains bore blaIMP and blaTEM genes on a single plasmid, as confirmed by trans...

    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Zhao WH,Hu ZQ,Chen G,Matsushita K,Fukuchi K,Shimamura T

    更新日期:2007-01-01 00:00:00

  • Multifunctionality and diversity of culturable bacterial communities strictly associated with spores of the plant beneficial symbiont Rhizophagus intraradices.

    abstract::Arbuscular Mycorrhizal Fungi (AMF) live in symbiosis with most crop plants and represent essential elements of soil fertility and plant nutrition and productivity, facilitating soil mineral nutrient uptake and protecting plants from biotic and abiotic stresses. These beneficial services may be mediated by the dense an...

    journal_title:Microbiological research

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    authors: Battini F,Cristani C,Giovannetti M,Agnolucci M

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Bellali S,Bou Khalil J,Fontanini A,Raoult D,Lagier JC

    更新日期:2020-06-01 00:00:00

  • Sensitive and rapid identification of Vibrio vulnificus by loop-mediated isothermal amplification.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Ren CH,Hu CQ,Luo P,Wang QB

    更新日期:2009-01-01 00:00:00

  • Purification and biochemical characterization of polygalacturonase II produced in semi-solid medium by a strain of Fusarium moniliforme.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Niture SK,Pant A

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  • Extracellular proteases from eight psychrotolerant Antarctic strains.

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    journal_title:Microbiological research

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    authors: Vazquez SC,Coria SH,MacCormack WP

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  • On the evolution of 'Indian Bison type' strains of Mycobacterium avium subspecies paratuberculosis.

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    journal_title:Microbiological research

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    authors: Sohal JS,Singh SV,Singh PK,Singh AV

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Wang KC,Huang CH,Chang PR,Huang MT,Fang SB

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  • The rad2 gene of haloarchaeum Halobacterium salinarum is functional in the repair of ultraviolet light induced DNA photoproducts.

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    pub_type: 杂志文章


    authors: Vafadarnejad E,Amoozgar MA,Khansha J,Fallahzade R

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Zheng B,Wang L,Liu L

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  • Role of σ-factor (orf21) in clavulanic acid production in Streptomyces clavuligerus NRRL3585.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Jnawali HN,Liou K,Sohng JK

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  • Use of metabolic and molecular methods for the identification of a Bacillus strain isolated from paper affected by foxing.

    abstract::Foxing of paper is a deterioration phenomenon occurring in the form of brown-yellowish spots, the abiotic and/or biotic causes of which are not yet completely understood. Nevertheless, microbiological infection has been recognized that may contribute to paper damage and therefore it becomes important to know the taxon...

    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: De Paolis MR,Lippi D

    更新日期:2008-01-01 00:00:00

  • Influence of microbial interactions and EPS/polysaccharide composition on nutrient removal activity in biofilms formed by strains found in wastewater treatment systems.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Andersson S,Dalhammar G,Kuttuva Rajarao G

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  • Trichoderma koningii as a biomineralizing fungous agent of calcium oxalate crystals in typical Argiudolls of the Los Padres Lake natural reserve (Buenos Aires, Argentina).

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    authors: Oyarbide F,Osterrieth ML,Cabello M

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  • Ethanol fermentation of mahula (Madhuca latifolia L.) flowers using free and immobilized yeast Saccharomyces cerevisiae.

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    authors: Swain MR,Kar S,Sahoo AK,Ray RC

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    authors: Meneses DP,Gudiña EJ,Fernandes F,Gonçalves LRB,Rodrigues LR,Rodrigues S

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    authors: Torres M,Romero M,Prado S,Dubert J,Tahrioui A,Otero A,Llamas I

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  • Construction of recombinant mercury resistant Acidithiobacillus caldus.

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    journal_title:Microbiological research

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    authors: Chen D,Lin J,Che Y,Liu X,Lin J

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  • Effects of compounds found in Nidus Vespae on the growth and cariogenic virulence factors of Streptococcus mutans.

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    authors: Guan X,Zhou Y,Liang X,Xiao J,He L,Li J

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  • In vivo gene expression and immunoreactivity of Leptospira collagenase.

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    journal_title:Microbiological research

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    authors: Janwitthayanan W,Keelawat S,Payungporn S,Lowanitchapat A,Suwancharoen D,Poovorawan Y,Chirathaworn C

    更新日期:2013-06-12 00:00:00

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Lebeau T,Jouenne T,Junter GA

    更新日期:2007-01-01 00:00:00

  • Isolation, purification, and identification of the virulence protein VirE2 from Agrobacterium tumefaciens.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Volokhina I,Sazonova I,Velikov V,Chumakov M

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  • Determination of laccase gene expression during degradation of 2,4,6-trinitrotoluene and its catabolic intermediates in Trametes versicolor.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Cheong S,Yeo S,Song HG,Choi HT

    更新日期:2006-01-01 00:00:00

  • Molecular cloning and homology modeling of protocatechuate 3,4-dioxygenase from Pseudomonas marginata.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Petersen E,Zuegg J,Ribbons DW,Schwab H

    更新日期:1996-12-01 00:00:00

  • Phenotypic and genotypic evaluation of slime production by conventional and molecular microbiological techniques.

    abstract::Twenty-nine staphylococcal isolates from different clinical samples were tested for slime production: phenotypic characterization was carried out using Christensen test (tube test) and Congo red agar plate test (CRA plate test), while the presence and expression of icaA and icaD genes were evaluated by real-time PCR. ...

    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Liberto MC,Matera G,Quirino A,Lamberti AG,Capicotto R,Puccio R,Barreca GS,Focà E,Cascio A,Focà A

    更新日期:2009-01-01 00:00:00

  • Bacillus amyloliquefaciens SB14 from rhizosphere alleviates Rhizoctonia damping-off disease on sugar beet.

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    journal_title:Microbiological research

    pub_type: 杂志文章


    authors: Karimi E,Safaie N,Shams-Baksh M,Mahmoudi B

    更新日期:2016-11-01 00:00:00

  • Current situation of biofuel production and its enhancement by CRISPR/Cas9-mediated genome engineering of microbial cells.

    abstract::Geopolitical and economic factors have motivated the scientific community to utilize renewable energy resources. In addition to the modifications in major steps and processes of biofuel production, manipulation of microbial genome engineering tools is essential in order to find sustainable solution of continuous deple...

    journal_title:Microbiological research

    pub_type: 杂志文章,评审


    authors: Javed MR,Noman M,Shahid M,Ahmed T,Khurshid M,Rashid MH,Ismail M,Sadaf M,Khan F

    更新日期:2019-02-01 00:00:00