Prof. Montarop Yamabhai

Year	Title	Journal
2012	Selection of single chain human monoclonal antibody (scFv) against Rabies virus by phage display technology Pruksametanan N., Yamabhai M., Khawplod P.	IEEE International Conference on Nano/Molecular Medicine and Engineering, NANOMED
Abstract: Human monoclonal antibodies against Rabies virus were selected from non-immunized human scFv library (YAMO-I library) and immunized library (Yamo-Rb library) by using phage display technology. The biopanning was performed for 2-5 rounds by using two types of inactivated rabies vaccines as targets. These are purified vero cell rabies vaccine (PVRV) and purified chick embryo cell vaccine (PCEC). A total of 14 positive clones from various method of biopanning that can bind to rabies, i.e.; IRA7c, IIIRC2c, IRC3c, IYCllc, IYCI2c, IYD1c, IYF5c, IIRD5v, IIYB5v, IIYG4v, IIYE5v, IIYG8v, IIYD4v and IVB4cv, were isolated and their genes were sequenced. The ELISA result showed that the positive clones always bind strongly to the targets that were used for biopanning; however some clones can cross-react to the related virus. These selected scFv antibodies will be tested for neutralization activities in vitro in the next step. © 2012 IEEE. Keyword: Human monoclonal antibody; Phage display; Rabies virus; Single-chain fragments(scFv) Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84880195559&doi=10.1109%2fNANOMED.2012.6509127&partnerID=40&md5=ff5168e5fb49be70a27d3563328875c9 DOI: 10.1109/NANOMED.2012.6509127
2012	Biomimicking the formation of nacre/shell: One step forward Khoushab F., Yamabhai M., Haller K.J.	Lecture Notes of the Institute for Computer Sciences, Social-Informatics and Telecommunications Engineering
Abstract: Chitin is biocompatible and biodegradable, with antimicrobial activity and low immunogenicity, and the second most abundant biopolymer after cellulose. It is one of the necessary components of the formation of nacre/shell. So harnessing behavior of chitin not only would lead to mimics of these structures but also to potential applications in academic research and industry. A peptide induces a unique chitin-based gel formation. Then, in the presence of CaCO3 a flexible structure was obtained. This structure and complex has many potential applications for different applications such as development of bioinspired instruments, or complexes, nanotechnology etc. It is early step toward mimicking the structure of nacre/shell. © 2012 ICST Institute for Computer Science, Social Informatics and Telecommunications Engineering. Keyword: Bioinspired; chitin; mimicking; nacre/shell; nanotechnology; peptide Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84869595514&doi=10.1007%2f978-3-642-32615-8_8&partnerID=40&md5=e1735f9ac0aa47ac88b80876e90ff4e5 DOI: 10.1007/978-3-642-32615-8_8
2012	Formation of chitin-based nanomaterials using a chitin-binding peptide selected by phage-display Khoushab F., Jaruseranee N., Tanthanuch W., Yamabhai M.	International Journal of Biological Macromolecules
Abstract: Targeting polymers with peptides is an efficient strategy to functionalize biomaterials. Phage display technology is one of the most powerful techniques for selecting specific peptides for a wide variety of targets. A method to select a chitin-binding peptide from a 12-mer random peptide library was successfully performed against chitin immobilized in wells of microtiter plates. The synthetic chitin binding peptide (ChiBP) could bind to chitin beads and disrupt their structure. This selected peptide was successfully used to immobilize alkaline phosphatase on chitin. In addition, the peptide could induce colloidal chitin in water to form a chitin coat on the surface of plastic tubes. Scanning electron microscopy (SEM) revealed that the peptide could induce colloidal chitin and chitohexaose to form networks when the temperature was raised to 42 °C. © 2012 Elsevier B.V. Keyword: Chitin; Nanobiotechnology; Nanomaterial; Peptide; Phage display Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84861132455&doi=10.1016%2fj.ijbiomac.2012.03.016&partnerID=40&md5=473aba37423d57494aae75538a5c2a3f DOI: 10.1016/j.ijbiomac.2012.03.016
2011	One-step detection of aflatoxin-B 1 using scFv-alkaline phosphatase-fusion selected from human phage display antibody library Rangnoi K., Jaruseranee N., O'Kennedy R., Pansri P., Yamabhai M.	Molecular Biotechnology
Abstract: A unique human phage display library was used to successfully generate a scFv to the highly carcinogenic toxin aflatoxin B1. Such an antibody has major potential applications in therapy and diagnostics. To further exploit its analytical capacity, the scFv was genetically fused to alkaline phosphatase, thereby generating a novel and highly sensitive self-indicating reagent. The performance of this reagent was further characterized, demonstrating its efficacy. The sensitivity of scFv-AP fusion was three-fold better than that of the scFv form. The ability of this human library to generate antibodies to a small hapten was clearly demonstrated and this is linked to its intrinsic diversity, which exceeds many existing conventional human libraries. Our results indicate that demography may influence the diversity of the repertoire of the library in terms of its capacity to generate antibodies to specific targets. Equally, the approach demonstrated should also be applicable for other haptens and larger antigens. © Springer Science+Business Media, LLC 2011. Keyword: Aflatoxin; Alkaline phosphatase; Antibody engineering; Competitive; ELISA; Hapten; Human; Phage display; Recombinant antibody; ScFv Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-80055018717&doi=10.1007%2fs12033-011-9398-2&partnerID=40&md5=53a576480854bd7817fcbf9d9e919802 DOI: 10.1007/s12033-011-9398-2
2011	Crystallization and preliminary crystallographic analysis of Β-mannanase from Bacillus licheniformis Songsiriritthigul C., Lapboonrueng S., Roytrakul S., Haltrich D., Yamabhai M.	Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Abstract: The mannan endo-1,4-Β-mannosidase (ManB) from Bacillus licheniformis strain DSM13 was overexpressed in Escherichia coli. Purification of the thermostable and alkali-stable recombinant mannanase yielded approximately 50 mg enzyme per litre of culture. Crystals were grown by hanging-drop vapour diffusion using a precipitant solution consisting of 12%(w/v) PEG 8000, 0.2 M magnesium acetate tetrahydrate and 0.1 M MES pH 6.5. The protein crystallized in the monoclinic space group P21, with two molecules per asymmetric unit and unit-cell parameters a = 48.58, b = 91.75, c = 89.55 Å, Β = 98.29°, and showed diffraction to 2.3 Å resolution. © 2011 International Union of Crystallography. All rights reserved. Keyword: Β-mannanase; Bacillus licheniformis strain DSM13; Glycosyl hydrolase family 26 Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-79951519964&doi=10.1107%2fS1744309110049067&partnerID=40&md5=e5fea6980212e4ef67386f2409b025e8 DOI: 10.1107/S1744309110049067
2011	Cloning, purification, and characterization of β-galactosidase from Bacillus licheniformis DSM 13 Juajun O., Nguyen T.-H., Maischberger T., Iqbal S., Haltrich D., Yamabhai M.	Applied Microbiology and Biotechnology
Abstract: The gene encoding homodimeric β-galactosidase (lacA) from Bacillus licheniformis DSM 13 was cloned and overexpressed in Escherichia coli, and the resulting recombinant enzyme was characterized in detail. The optimum temperature and pH of the enzyme, for both o-nitrophenyl-β-d-galactoside (oNPG) and lactose hydrolysis, were 50°C and 6.5, respectively. The recombinant enzyme is stable in the range of pH 5 to 9 at 37°C and over a wide range of temperatures (4-42°C) at pH 6.5 for up to 1 month. The K m values of LacA for lactose and oNPG are 169 and 13.7 mM, respectively, and it is strongly inhibited by the hydrolysis products, i.e., glucose and galactose. The monovalent ions Na+ and K+ in the concentration range of 1-100 mM as well as the divalent metal cations Mg2+, Mn2+, and Ca2+ at a concentration of 1 mM slightly activate enzyme activity. This enzyme can be beneficial for application in lactose hydrolysis especially at elevated temperatures due to its pronounced temperature stability; however, the transgalactosylation potential of this enzyme for the production of galacto-oligosaccharides (GOS) from lactose was low, with only 12% GOS (w/w) of total sugars obtained when the initial lactose concentration was 200 g/L. © 2010 Springer-Verlag. Keyword: β-Galactosidase; Bacillus licheniformis; Lactose hydrolysis; Recombinant; Transglycosylation Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-79251595403&doi=10.1007%2fs00253-010-2862-2&partnerID=40&md5=0330ed08ee9a7c99cda85df55d983e95 DOI: 10.1007/s00253-010-2862-2
2011	Efficient e. coli expression systems for the production of recombinant β-mannanases and other bacterial extracellular enzymes Yamabhai M., Buranabanyat B., Jaruseranee N., Songsiriritthigul C.	Bioengineered Bugs
Abstract: Two Escherichia coli expression systems based on T7 RNA polymerase promoter (pET system) and tac promoter (pFLAG system) have been used for the production and secretion of recombinant β-mannanases from Bacillus sp. Both E. coli OmpA signal peptide and native Bacillus signal peptide could be used efficiently for the secretion of recombinant enzymes into periplasmic space and culture media. The genes could be induced for overexpression with 0.1-1 mM isopropyl-β-D-1-thiogalactopyranoside (IPTG) when the OD600 of the culture broth reached 0.6-1.5. The recombinant enzymes could be harvested from whole cell lysate, perimplasmic extract or culture broth after induction for 4-20 hours. Since the enzyme is C-terminally tagged with hexahistidine, the recombinant enzymes could be conveniently purified to apparent homogeneity by onestep immobilized-metal affinity chromatography (IMAC) using Ni-NTA resins. The characteristics of purified recombinant β-mannanases from B. licheniformis and B. subtilis, which share 78% amino acid identity, are slightly different. These systems should be applicable for the production of various recombinant bacterial extracellular enzymes. © 2011 Landes Bioscience. Keyword: Bacteria; Enzyme; Escherichia coli; Expression; Extracellular; Mannanase; OmpA; Recombinant; T7 promoter; Tac promoter Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-78751673067&doi=10.4161%2fbbug.2.1.13419&partnerID=40&md5=5008ba3d8436f5adf34325662279ed46 DOI: 10.4161/bbug.2.1.13419
2010	β-galactosidase from Lactobacillus pentosus: Purification, characterization and formation of galacto-oligosaccharides Maischberger T., Leitner E., Nitisinprasert S., Juajun O., Yamabhai M., Nguyen T.-H., Haltrich D.	Biotechnology Journal
Abstract: A novel heterodimeric β-galactosidase with a molecular mass of 105 kDa was purified from crude cell extracts of the soil isolate Lactobacillus pentosus KUB-ST10-1 using ammonium sulphate fractionation followed by hydrophobic interaction and affinity chromatography. The electrophoretically homogenous enzyme has a specific activity of 97 UoNPG/mg protein. The Km, kcat and kcat/Km values for lactose and o-nitrophenyl-β-D-galactopyranoside (oNPG) were 38 mM, 20 s-1, 530 M-1•s-1 and 1.67 mM, 540 s -1, 325 000 M-1•s-1, respectively. The temperature optimum of β-galactosidase activity was 60-65° C for a 10-min assay, which is considerably higher than the values reported for other lactobacillal β-galactosidases. Mg2+ ions enhanced both activity and stability significantly. L. pentosus β-galactosidase was used for the production of prebiotic galactooligosaccharides (GOS) from lactose. A maximum yield of 31% GOS of total sugars was obtained at 78% lactose conversion. The enzyme showed a strong preference for the formation of β-(1→3) and β-(1→6) linkages, and the main transgalactosylation products identified were the disaccharides β-D-Galp-(1→6)-D-Glc, β-D-Galp-(1→3)-D-Glc, β-D-Galp-(1→6)-D-Gal, β-D-Galp-(1→3)-DGal, and the trisaccharides β-D-Galp-(1→3)- D-Lac, β-D-Galp-(1→6)-D-Lac. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA. Keyword: Biocatalysis; Food biotechnology; Lactase; Lactic acid bacteria; Transgalactosylation Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-77955637291&doi=10.1002%2fbiot.201000126&partnerID=40&md5=2af2ceff09816772df192ec34f1b3c83 DOI: 10.1002/biot.201000126
2010	Characterisation of recombinant pyranose oxidase from the cultivated mycorrhizal basidiomycete Lyophyllum shimeji (hon-shimeji) Salaheddin C., Takakura Y., Tsunashima M., Stranzinger B., Spadiut O., Yamabhai M., Peterbauer C.K., Haltrich D.	Microbial Cell Factories
Abstract: Background: The flavin-dependent enzyme pyranose 2-oxidase (P2Ox) has gained increased attention during the last years because of a number of attractive applications for this enzyme. P2Ox is a unique biocatalyst with high potential for biotransformations of carbohydrates and in synthetic carbohydrate chemistry. Recently, it was shown that P2Ox is useful as bioelement in biofuel cells, replacing glucose oxidase (GOx), which traditionally is used in these applications. P2Ox offers several advantages over GOx for this application, e.g., its much broader substrate specificity. Because of this renewed interest in P2Ox, knowledge on novel pyranose oxidases isolated from organisms other than white-rot fungi, which represent the traditional source of this enzyme, is of importance, as these novel enzymes might differ in their biochemical and physical properties.Results: We isolated and over-expressed the p2ox gene encoding P2Ox from the ectomycorrhizal fungus Lyophyllum shimeji. The p2ox cDNA was inserted into the bacterial expression vector pET21a(+) and successfully expressed in E. coli Rosetta 2. We obtained active, flavinylated recombinant P2Ox in yields of approximately 130 mg per L of medium. The enzyme was purified by a two-step procedure based on anion exchange chromatography and preparative native PAGE, yielding an apparently homogenous enzyme preparation with a specific activity of 1.92 U/mg (using glucose and air oxygen as the substrates). Recombinant P2Ox from L. shimeji was characterized in some detail with respect to its physical and catalytic properties, and compared to the well-characterised enzymes from Phanerochaete chrysosporium and Trametes multicolor.Conclusion: L. shimeji P2Ox shows properties that are comparable to those of P2Ox from white-rot fungal origin, and is in general characterised by lower Kmand kcatvalues both for electron donor (sugar) as well as electron acceptor (ferrocenium ion, 1,4-benzoquinone, 2,6-dichloroindophenol). While L. shimeji P2Ox is the least thermostable of these three enzymes (melting temperature Tmof 54.9°C; half-life time of activity τ1/2of 0.12 at 50°C and pH 6.5), P. chrysosporium P2Ox showed remarkable thermostability with Tmof 75.4°C and τ1/2of 96 h under identical conditions. © 2010 Salaheddin et al; licensee BioMed Central Ltd. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-77954487790&doi=10.1186%2f1475-2859-9-57&partnerID=40&md5=3b07cb1c528eef969d60dd6b85c5e618 DOI: 10.1186/1475-2859-9-57
2010	Expression and characterization of Bacillus licheniformis chitinase (ChiA), suitable for bioconversion of chitin waste Songsiriritthigul C., Lapboonrueng S., Pechsrichuang P., Pesatcha P., Yamabhai M.	Bioresource Technology
Abstract: Chitinase (EC 3.2.1.14) is an enzyme with multiple industrial applications. These include bioconversion of chitin waste, a highly resistant and abundant biopolymer from crustacean food industry, into glucosamine and chito-oligosaccharide value-added products. This paper reports on the expression of endochitinase (ChiA) from Bacillus licheniformis strain DSM8785 in E. coli and characterization of the recombinant enzyme. Recombinant ChiA could efficiently convert colloidal chitin to N-acetyl glucosamine and chitobiose at pH 4.0, 6.0 and 9.0 at 50 °C and retained its activity up to 3 days under these conditions, suggesting that this enzyme is suitable for bioconversion of chitin waste. © 2010 Elsevier Ltd. All rights reserved. Keyword: Bacillus licheniformis; Bioconversion; Chitin; Chitinase; Waste Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-76749113742&doi=10.1016%2fj.biortech.2010.01.036&partnerID=40&md5=d6198150e2e2e4691c2412b46ea0a75d DOI: 10.1016/j.biortech.2010.01.036