Abstract:
:Laccases are polyphenol oxidoreductases used in a number of industrial applications. Due to the increasing demand for these "green catalysis" enzymes, the identification and biochemical characterisation of their novel properties is essential. In our study, cloned Madurella mycetomatis laccase (mmlac) genes were heterologously expressed in the methylotrophic yeast host Pichia pastoris. The high yield of the active recombinant protein in P. pastoris demonstrates the efficiency of a reliably constructed plasmid to express the laccase gene. The optimal biochemical conditions for the successfully expressed MmLac enzyme were identified. Detailed structural properties of the recombinant laccase were determined, and its utility in decolourisation and textile bleaching applications was examined. MmLac demonstrates good activity in an acidic pH range (4.0-6.0); is stable in the presence of cationic metals, organic solvents and under high temperatures (50-60 °C); and is stable for long-term storage at - 20 °C and - 80 °C for up to eight weeks. The structural analysis revealed that the catalytic residues are partially similar to other laccases. MmLac resulted in an increase in whiteness, whilst demonstrating high efficiency and stability and requiring the input of fewer chemicals. The performance of this enzyme makes it worthy of investigation for use in textile biotechnology applications, as well as within environmental and food technologies.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Tülek A,Karataş E,Çakar MM,Aydın D,Yılmazcan Ö,Binay Bdoi
10.1007/s12033-020-00281-9subject
Has Abstractpub_date
2021-01-01 00:00:00pages
24-39issue
1eissn
1073-6085issn
1559-0305pii
10.1007/s12033-020-00281-9journal_volume
63pub_type
杂志文章abstract::A shuttle vector pZL1 which can replicate both in Gluconobacter oxydans and Escherichia coli was constructed based on G. oxydans DSM2003 cryptic plasmid pGOX3, a homology of G. oxydans 621H pGOX3, and E. coli cloning vector pUC18. It was found to be stably maintained in G. oxydans during the serial subcultures in the ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9293-2
更新日期:2010-11-01 00:00:00
abstract::The recombinant acidstable α-amylase (Ba-amy) of acidophilic bacterium Bacillus acidicola TSAS1 has been produced extracellularly using a combination of cloning (E. coli and P. pastoris) and physico-chemical treatment strategies. A total of 150,000 U/L of Ba-amy were attained under constitutive promoter in P. pastoris...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-016-9970-x
更新日期:2016-11-01 00:00:00
abstract::Adenoviral dodecahedron (Dd) is a virus-like particle composed of twelve pentameric penton base (Pb) proteins, responsible for adenovirus cell penetration. It is generated spontaneously in the baculovirus system upon expression of the Pb gene of adenovirus serotype 3. This particle shows remarkable cell penetration ab...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-015-9850-9
更新日期:2015-06-01 00:00:00
abstract::Human adipose tissue-derived stromal cells (ADSCs) are being evaluated for cardiovascular repair. We developed an ex vivo method for producing angiogenic ADSCs transduced with a self-inactivating lentiviral vector (LV) expressing the enhanced green fluorescence protein (EGFP) from an internal cytomegalovirus (CMV) pro...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9537-4
更新日期:2013-05-01 00:00:00
abstract::The induction of several secondary metabolites in plants is one of the most commonly observed effects after the external addition of methyl jasmonate (MeJA). After the elicitation of Catharanthus roseus hairy roots with different concentrations of MeJA, changes in the accumulation of alkaloids such as ajmalicine, serp...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9111-2
更新日期:2009-03-01 00:00:00
abstract::Previous studies have shown that mRNA-electroporated dendritic cells (DCs) are able to process and present tumor-associated antigens, leading to the activation of tumor-specific T cells in vitro and in vivo. However, the optimal maturation state of antigen loading and half-life of the mRNA-translated protein product a...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9071-6
更新日期:2008-10-01 00:00:00
abstract::RNA interference (RNAi) efficiently induces sequence-specific gene silencing in mammalian cells through short interfering RNA (siRNA) of 21-23 nucleotides synthesized in vitro or expressed by DNA-based vector. However, introduction of siRNA into mammalian cells by transfection limits the application of RNAi, especiall...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:28:3:195
更新日期:2004-11-01 00:00:00
abstract::The significance of actin cytoskeleton on cell growth was historically studied using toxic drugs, such as cytochalasin. However, it is possible that unpredictable effects of these agents may have influenced the reported observations. In our study, we have established a drug-free system using cofilin overexpression to ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:31:1:001
更新日期:2005-09-01 00:00:00
abstract::Tea, a beverage crop, is a rich source of polyphenols and polysaccharides which greatly attribute to its importance. However, oxidation and precipitation of these compounds during nucleic acids extraction is a limitation to molecular biology and genomic studies. On isolation of total RNA from root tissue using establi...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9476-5
更新日期:2012-09-01 00:00:00
abstract::Isolation of RNA from recalcitrant tree tissues has been problematic due to large amounts of secondary metabolites and interfering compounds in their cells. We have developed an efficient RNA extraction method, which yielded high-quality RNA preparations from tissues of the lychee tree. The method reported here utiliz...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0073-6
更新日期:2008-01-01 00:00:00
abstract::This work describes protocols for the production of single-chain antibody and T-cell receptor fragments in E. coli. A choice of methods is given for the purification of the recombinant fragments that rely on the use of either immunoaffinity or metal chelate affinity chromatography. The TCR fragments may have to be den...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02779017
更新日期:1995-12-01 00:00:00
abstract::Electroporation of cells in the presence of DNA is widely used for the introduction of transgenes either stably or transiently into bacterial, fungal, animal, and plant cells. A review of the literature shows that electroporation parameters are often reported in an incomplete or incorrect manner, forcing researchers t...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02821542
更新日期:1997-02-01 00:00:00
abstract::Papaya ringspot virus (PRSV) causes severe economic losses in both cucurbits and papaya throughout the tropics and subtropics. Development of PRSV-resistant transgenic plants faces a major hurdle in achieving resistance against geographically distinct isolates. One of the major reasons of failing to achieve the broad-...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-009-9205-5
更新日期:2010-01-01 00:00:00
abstract::An inexpensive monophasic reagent has been developed for the extraction of total RNA from cells or tissues. The main ingredients of the reagent are Phenol, Isoamyl alcohol, Guanidinium isothiocyanate, and Beta-mercaptoethanol (PIG-B). The quality and yield of RNA obtained by this reagent is at par with that obtained b...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02752699
更新日期:1998-02-01 00:00:00
abstract::The low economic profits of feather recycling lead that the large amount of feathers is currently discarded in China. To convert feather hydrolysates into GSH with high values, surface display of the bifunctional glutathione synthetase encoded by gcsgs from Streptococcus thermophilus on Saccharomyces cerevisiae and th...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-014-9750-4
更新日期:2014-08-01 00:00:00
abstract::Since it was first introduced in late 1990s Wave bioreactor has been used for protein production by mammalian and insect cell lines. However, using Wave bioreactor to produce human monoclonal antibody by stable Drosophila Schneider 2 (S2) cell transfectants has not been reported before. In this study, S2 cells were co...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9484-5
更新日期:2012-10-01 00:00:00
abstract::Yeast artificial chromosome (YAC) cloning systems enable the cloning of DNA stretches of 50 to well over 2000 kb. This makes it possible to study large intact regions of DNA in detail, by restriction mapping the YAC to produce a physical map and by examining the YAC for coding sequences or genes. YACs are important fo...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02921558
更新日期:1994-04-01 00:00:00
abstract::Pyrococcus woesei (DSM 3773) beta-galactosidase gene amplified by polymerase chain reaction was cloned into KpnI and HindIII binding sites of pET-30LIC expression plasmid. The obtained pGal2 (6785 bp) transcription vector was then transferred to Escherichia coli B121 (DE3) cells. High identity (99.9%) of DNA sequences...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02740841
更新日期:1998-12-01 00:00:00
abstract::The macrofungus Cordyceps militaris contains many kinds of bioactive ingredients that are regulated by functional genes, but the functions of many genes in C. militaris are still unknown. In this study, to improve the frequency of homologous integration, a genetic transformation system based on a split-marker approach...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0080-9
更新日期:2018-05-01 00:00:00
abstract::The MYB proteins constitute one of the largest transcription factor families in plants. Much research has been performed to determine their structures, functions, and evolution, especially in the model plants, Arabidopsis, and rice. However, this transcription factor family has been much less studied in wheat (Triticu...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9486-3
更新日期:2012-10-01 00:00:00
abstract::Addition of taxol into suspension cultures of Taxus cuspidata induced cell apoptosis, which was confirmed by gel electrophoresis of the DNA ladders indicating the progressive delineation of fragmented nuclear DNA (nDNA) into distinct bodies. The additional taxol not only changed the microtubule assembly of cells, but ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:20:2:137
更新日期:2002-02-01 00:00:00
abstract::ATP-binding cassette transporter G2 (ABCG2) gene encodes a protein that has a wide variety of substrates and is responsible for the active secretion of clinically and toxicologically important molecules into milk. Although known in many species, this marks the first time this gene product has been reported in goats. I...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-9024-5
更新日期:2008-05-01 00:00:00
abstract::Small ubiquitin-related modifier (SUMO) fusion system has been shown to be efficient for enhancing expression and preventing degradation of the target protein. We showed herein that SUMO fusion to human keratinocyte growth factor 2 (hKGF-2) gene was feasible and it significantly enhanced protein expression and its eff...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9135-7
更新日期:2009-05-01 00:00:00
abstract::The polyamines spermine, spermidine, and putrescine are small organic molecules one or more of which are present in all living organisms. Many natural products contain polyamine residues. Polyamines are synthesized by a highly regulated pathway from arginine or ornithine and also can be transported in and out of cells...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02788682
更新日期:1999-06-01 00:00:00
abstract::NK cells have been seen as potential agents in adoptive immunotherapy for cancer. The main challenge for the success of this approach is to obtain a great quantity of activated NK cells for adoptive transfer. The present study had aimed to evaluate the effect of a feeder layer of irradiated MSCs in the in vitro expans...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-020-00250-2
更新日期:2020-05-01 00:00:00
abstract::Agrobacterium-mediated transformation is the method of choice to engineer desirable genes into plants. Here we describe a protocol for demonstrating T-DNA transfer from Agrobacterium into the economically important graminaceous plant maize. Expression of the T-DNA-located GUS gene was observed with high efficiency on ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:13:2:165
更新日期:1999-12-01 00:00:00
abstract::A sensitive method for staining proteins after transfer from polyacrylamide gels to nitrocellulose paper is described. Transferred proteins are first derivatized by reaction of the nitrocellulose replica with sulfosuccinimidobiotin and are then reacted sequentially with streptavidin, rabbit anti-streptavidin, and hors...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:15:2:161
更新日期:2000-06-01 00:00:00
abstract::Recombinant proteins are useful tools in biological research, drug development, and drug screening. Specially designed expression vectors have been developed to introduce cDNA for recombinant protein expression in mammalian cells. We have combined a dicistronic mRNA design for expression of the recombinant protein, us...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02745860
更新日期:1998-08-01 00:00:00
abstract::RNA editing is one of the post-transcriptional RNA processes. RNA editing generates RNA and protein diversity in eukaryotes and results in specific amino acid substitutions, deletions, and changes in gene expression levels. Adenosine-to-inosine RNA editing represents the most important class of editing in human and af...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-012-9498-7
更新日期:2012-09-01 00:00:00
abstract::Enzymes are key molecules in signal-transduction pathways. However, only a small fraction of more than 500 human kinases, 300 human proteases and 200 human phosphatases is characterised so far. Peptide microarray based technologies for extremely efficient profiling of enzyme substrate specificity emerged in the last y...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-011-9402-x
更新日期:2011-11-01 00:00:00