Abstract:
OBJECTIVES:L-isoleucine dioxygenase (IDO) specifically transforms L-isoleucine (Ile) to 4-hydroxyisoleucine (4-HIL), and 4-HIL is a promising drug for diabetes. To enhance the activity and catalytic efficiency of IDO, we used directed evolution and site-specific mutagenesis. RESULTS:The IDO gene (ido) derived from Bacillus weihenstephanensis was cloned and expressed in Escherichia coli. Directed evolution using error prone (EP)-PCR and site-specific mutagenesis were conducted. Two improved mutants were obtained after one round of EP-PCR, with IdoN126H exhibiting a 2.8-fold increase in activity. Two improved mutants were obtained through site-specific mutagenesis, with IdoT130K showing a 170% increase in activity. Although the activity of the combined mutant IdoN126H/T130K (0.95 ± 0.08 U/mg) was slightly higher than that of the wild-type Ido, its catalytic efficiency was 2.4-fold and 3.0-fold higher than Ido with Ile and α-ketoglutaric acid as substrates. After biotransformation of Ile by E. coli BL21(DE3) expressing IdoN126H/T130K and Ido, 66.50 ± 0.99 mM and 26.09 ± 1.85 mM 4-HIL was synthesized, respectively, in 24 h. CONCLUSION:IdoN126H/T130K had a higher enzyme activity and catalytic efficiency and can therefore be used as a more suitable candidate for 4-HIL production.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Huang S,Shi Fdoi
10.1007/s10529-018-2566-8subject
Has Abstractpub_date
2018-08-01 00:00:00pages
1227-1235issue
8eissn
0141-5492issn
1573-6776pii
10.1007/s10529-018-2566-8journal_volume
40pub_type
杂志文章abstract::The gene for beta-agarase of an Agarivorans sp. JA-1 was expressed in Bacillus subtilis strain DB104 for efficient and economical mass-production of the enzyme. We isolated 360 mg protein with a specific activity of 201 U/mg from the culture broth. The efficiency of production was approximately 130-fold higher than th...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9634-4
更新日期:2008-05-01 00:00:00
abstract::Pectinase was immobilized on a sodium alginate support using glutaraldehyde and retained 66% activity. The optimal pH for activity shifted from 3.0 to 3.5 after immobilization; however, the optimum temperature remained unchanged at 40 degrees C. The immobilized enzyme also had a higher thermal stability and reusabilit...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-007-9409-3
更新日期:2007-09-01 00:00:00
abstract:OBJECTIVES:Establish a method to indirectly measure evaporation in microwell-based cell culture systems and show that the proposed method allows compensating for liquid losses in fed-batch processes. RESULTS:A correlation between evaporation and the concentration of Na+ was found (R2 = 0.95) when using the 24-well-bas...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-018-2556-x
更新日期:2018-07-01 00:00:00
abstract::The promoter region of the pur operon, which contains 12 genes for inosine monophosphate biosynthesis from phosphoribosylpyrophosphate, and the purA gene, encoding the adenylosuccinate synthetase, were compared among wild-type and three purine-producing Bacillus subtilis strains. A single nucleotide deletion at positi...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-006-9020-z
更新日期:2006-06-01 00:00:00
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
更新日期:2004-10-01 00:00:00
abstract::A recombinant allophycocyanin (rAPC), used for treatment of tumors, has been expressed in E. coli which was grown in glucose fed-batch culture in a 30 l fermentor. Recombinant allophycocyanin was purified from soluble E. coli cell lysate using hydrophobic interaction chromatography followed by chromatography using amy...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-5794-7
更新日期:2005-06-01 00:00:00
abstract:OBJECTIVES:Regenerative medicine approaches using reprogrammed or transdifferentiated cells require efficient single cell expression profiling to analyze culture homogeneity for quality control and recipients' safety. RESULTS:While antigen-antibody based systems have been developed for several proteins, probing at the...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-015-1833-1
更新日期:2015-08-01 00:00:00
abstract::The peptide, Ala-Pro-Ala-Arg (APAR), was selected from the screening of a tetrapeptide combinatorial synthetic library as the ligand for affinity purification of an anti-Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) monoclonal antibody (Mab) developed in mouse ascitis. The affinity chromatographic matrix o...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/a:1025464918453
更新日期:2003-09-01 00:00:00
abstract::Aptamers against inactive Vibrio alginolyticus were selected from an 82-nt ssDNA random library by systematic evolution of ligands by exponential enrichment. After 15 rounds of selection, the final pool of aptamers was highly specific for inactivated V. alginolyticus and had a dissociation constant of 27.5 ± 9.2 nM. U...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-013-1154-1
更新日期:2013-06-01 00:00:00
abstract:OBJECTIVE:To construct effective genetic expression parts controlling transcription and translation initiation for synthetic biology and heterologous expression in Corynebacterium glutamicum. RESULT:Twelve highly expressed genes were identified from the proteomic data of C. glutamicum. Their related sequences were use...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-016-2196-y
更新日期:2016-12-01 00:00:00
abstract::The biomacromolecule, gelatin, has increasingly been used in biomedicine-beyond its traditional use in food and cosmetics. The appealing advantages of gelatin, such as its cell-adhesive structure, low cost, off-the-shelf availability, high biocompatibility, biodegradability and low immunogenicity, among others, have m...
journal_title:Biotechnology letters
pub_type: 杂志文章,评审
doi:10.1007/s10529-015-1907-0
更新日期:2015-11-01 00:00:00
abstract::Marinomonas sp. NJ522, isolated from Antarctic sea ice, produces a cold-active iron superoxide dismutase (SOD; EC 1.15.1.1). The purified SOD was dimeric and had an approx. Mr of 48 kDa. Highest activity was detected from pH 8 to 10 and at 40 degrees C (assayed over 10 min). Activity at 0 degree C was nearly 35% of th...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-4951-3
更新日期:2006-01-01 00:00:00
abstract::Baker's yeast suspensions were incubated at different pressures (from 1 bar to 6 bar) and different gases [air, O(2) and a mixture of 8% (v/v) CO(2), 21% O(2) and N(2)]. Raising the air pressure from 1 bar to 6 bar stimulated cell growth but had no effect on leavening ability or viability of the cells. A 50% reduction...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/B:BILE.0000036604.20963.da
更新日期:2004-08-01 00:00:00
abstract::A single-compartmented microbial fuel cell composed of a graphite felt anode modified with Neutral Red (NR-anode) and a porous Fe(II)-carbon cathode (FeC-cathode) were compared for electricity generation from Microbacterium sp. and Pseudomonas sp. under identical conditions. Pseudomonas sp. was more than four times th...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-009-0184-1
更新日期:2010-04-01 00:00:00
abstract::Hydrophobin-fused domain III of dengue envelope proteins serotypes 1 and 2 were expressed in Rachiplusia nu larvae and purified by aqueous two-phase system. This biotechnological approach of hydrophobin-fused proteins, which allowed obtaining 97.7 µg/larva of fusion protein DomIII serotype 1 and 61.4 µg/larva of fusio...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-019-02767-2
更新日期:2020-03-01 00:00:00
abstract::L-threo-3,4-Dihydroxyphenylserine (DOPS) is a chiral unnatural beta-hydroxy amino acid used for the treatment of Parkinson disease. We developed a continuous bioconversion system for DOPS production that uses whole-cell biocatalyst of recombinant Escherichia coli expressing L-threonine aldolase (L-TA) genes cloned fro...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9885-0
更新日期:2009-03-01 00:00:00
abstract::The penicillin G amidase (PGA) from Alcaligenes faecalis, which has interesting properties for use in combinatorial biochemistry, was produced by recombinant expression in Escherichia coli. The corresponding gene was cloned into a multicopy vector under the strict regulatory control of the rhamnose inducible promoter....
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/a:1022498314354
更新日期:2003-03-01 00:00:00
abstract::Tandem affinity purification (TAP) is a methodology for the isolation of protein complexes from endogenous sources. It involves incorporation of a dual-affinity tag into the protein of interest and introduction of the construct into desired cell lines or organisms. Using the two affinity handles, the protein complex a...
journal_title:Biotechnology letters
pub_type: 杂志文章,评审
doi:10.1007/s10529-011-0592-x
更新日期:2011-08-01 00:00:00
abstract::Bacterial and fungal contaminants of enset (Ensete ventricosum) cultures and microbes associated with surface-sterilized field material were identified by 16S/26S rDNA sequencing. Ten bacterial species were identified in 16 isolates from in vitro cultures and seven in 10 isolates from field clones. Three yeast species...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-004-6032-4
更新日期:2004-12-01 00:00:00
abstract::A carboxylesterase gene from thermophilic bacterium, Alicyclobacillus tengchongensis, was cloned and expressed in Escherichia coli BL21 (DE3). The gene coded for a 513 amino acid protein with a calculated molecular mass of 57.82 kDa. The deduced amino acid sequence had structural features highly conserved among serine...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-013-1195-5
更新日期:2013-08-01 00:00:00
abstract::A sigma 70-like gene, sigA, has been identified from Corynebacterium ammoniagenes. The sigA gene encodes a polypeptide of 467 amino acids with a calculated molecular mass of 52036 Da. The deduced amino acid sequence preserves the common motifs of the primary sigma factors and shows very high similarity to those of Sig...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000021962.16204.08
更新日期:2004-04-01 00:00:00
abstract::The Bacillus pumilus SG2 chitinase gene (ChiS) and its truncated form lacking chitin binding (ChBD) and fibronectin type III (FnIII) domains were transformed to Arabidopsis plants and the expression, functionality and antifungal activity of the recombinant proteins were investigated. Results showed that while the two ...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-009-0192-1
更新日期:2010-04-01 00:00:00
abstract::Alginate production by Azotobacter vinelandii growing in chemostat cultures was evaluated under different O(2) transfer rates (OTR). As a result of modifying the culture's agitation rate from 300 to 500 rpm, the OTR increased from 9 to 15.1 mmol l(-1) h(-1) and a slight variation in the alginate production (1.7-2.2 g ...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-009-9949-9
更新日期:2009-06-01 00:00:00
abstract::The ability of Aspergillus strains to utilize thin stillage to produce malic acid was compared. The highest malic acid was produced by Aspergillus niger ATCC 9142 at 17 g l(-1). Biomass production from thin stillage was similar with all strains but ATCC 10577 was the highest at 19 g l(-1). The highest malic acid yield...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-011-0720-7
更新日期:2011-12-01 00:00:00
abstract::Phanerochaete chrysoporium morphology and manganese peroxidase (MnP) productivity was characterised in a scalable, modularised 1145 cm(3) membrane gradostat reactor in response to switching between an enhanced production medium and a nutrient limited feed (50% C and N reduction). Irrespective of the feed composition u...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-010-0443-1
更新日期:2011-02-01 00:00:00
abstract::Fermentation kinetics of Penicillium aculeatum ATCC 10409 demonstrated that fungal growth and dextranase release are decoupled. Inoculation by conidia or mycelia resulted in identical kinetics. Two new isoenzymes of the dextranase were characterized regarding their kinetic constants, pI, MW, activation energy and stab...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9659-8
更新日期:2008-06-01 00:00:00
abstract::Constant medium feeding rate and intermittent fed-batch fermentation strategies were investigated aiming to increase the yields of γ-decalactone production by Yarrowia lipolytica, using methyl ricinoleate as substrate and ricinoleic acid source. The accumulation of another compound, 3-hydroxy-γ-decalactone, was also a...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-011-0824-0
更新日期:2012-04-01 00:00:00
abstract::The biotin-binding capacity in the wells of a streptavidin-coated PCR plate were quantified by means of a fluorescence intensity assay utilising biotin-labelled fluorescein and a colorimetric assay using biotin-labelled alkaline phosphatase. The biotin binding capacities were determined to be 59 and 58 pmol respective...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/a:1024951322955
更新日期:2003-08-01 00:00:00
abstract::Electrotransformation of Thermoanaerobacter ethanolicus JW200 was achieved using the plasmid, pTE16, and a pUC-based suicide vector, pTEA2. The construct pTE16 is based on the Escherichia coli-Clostridium perfringens shuttle vector pJIR715 and contains a thermostable chloramphenicol (Cm) resistance cassette. Evidence ...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-006-9184-6
更新日期:2006-12-01 00:00:00
abstract::A brief historical perspective on metagenomics is given followed by a discussion of the rapid progress in this field largely defined by transition to the next generation sequencing technologies. Problems and challenges connected to this transition are also addressed. The review focuses on recent literature describing ...
journal_title:Biotechnology letters
pub_type: 历史文章,杂志文章,评审
doi:10.1007/s10529-010-0306-9
更新日期:2010-10-01 00:00:00