VAAM-Jahrestagung 2012 18.–21. März in Tübingen

13816S rRNA genes was applied to acquire an overview about composition ofmicrobial communities in both types of digesters. Hydrolytic microbialcommunities were characterized applying a functional oligonucleotidemicroarray targeting 756 different cellulase genes.The pyrosequencing results showed that members of the phylumFirmicutes dominated in both digesters ranging from 73.8-84.5%.Representatives of the phylum Actinobacteria were the second abundantgroup found in both digesters, with a lower proportion detected in thethermophilic one. Significant differences in microbial communitystructures of the two digesters were found at finer taxonomical levels. Thefunctional oligonucleotide microarray focuses on the detection of fourdifferent cellulase gene families and results revealed similar functionaldistribution patterns between the two types of digesters although a higherdiversity within the cellulolytic microbial community was found inmesophilic digesters.In conclusion, this study could show that the operating conditions didaffect the diversity of microbial community, while the effect on thehydrolytic bacterial communities involved seems to be less pronounced.OTP001Molecular cloning of enantioselective ester hydrolase fromBacillus pumilusV. VermaShri Mata Vaishno Devi University , Biotechnology, Katra, IndiaA gene from Bacillus pumilus expressed under its native promoter wascloned in Escherichia coli. Recombinant B. pumilus esterase (BPE) affectsthe kinetic resolution of racemic mixtures such as unsubstituted andsubstituted 1-(phenyl)ethanols (E ~ 33–103), ethyl 3-hydroxy-3-phenylpropanoate (E ~ 45–71), trans-4-fluorophenyl-3-hydroxymethyl-Nmethylpiperidine(E ~ 10–13) and ethyl 2- hydroxy-4-phenylbutyrate (E~7). The enzyme is composed of a 34-amino acid signal peptide and a 181-amino acid mature protein corresponding to a molecular weight of ~19.2kD and pI ~ 9.4. 3-D the structural model of the enzyme built byhomology modelling using the atomic coordinates from the crystalstructure of B. subtilis lipase (LipA) showed a compact minimal a/bhydrolase fold.Biography: Prof. V. Verma, a former scientist of Indian Institute of IntegrativeMedicine (formerly known as Reg. Res. Lab), Jammu , a CSIR researchlaboratory and presently Professor of Biotechnology, Shri Mata Vaishno DeviUniversity, Katra (J&K) is acclaimed for his work in microbial biotechnology.His work in this field mainly related to the cloning & heterlogous overexpresionof microbial genes encoding enantio-specific enzymes known forresolving the racemic drug intermediates. Besides, he did pioneering work inthe development of fermentation based technologies for the mass production ofselected microbial isolates as biocontrol agents as part of integrated nutrient &disease management of agriculturally important plants. His research areasinclude microbial gene cloning & their heterologous expression, fermentationtechnology for organic agriculture and DNA finger printing of the importantmicrobial isolates for IPR & registration purposes.Prof. Verma is recipient of anumber of national/international awards in Biological Sciences andBiotechnology. He is Fellow of a number of Academies of Sciences in India.Recently he received INDUSTRIAL MEDAL AWARD from the BiotechResearch Society of India for his outstanding contributions in Biotechnology.OTP002Isolation and characterization of novel potent Cr (VI)reducing alkaliphilic bacterim from hypersaline soda lakesA. Ibrahim*, M. El-Tayeb, Y. El-Badawi, A. Al-SalamahBotany and Microbiology, Riyadh, Saudi ArabiaA strain KSUCr3 with extremely high Cr(VI)-reducing ability underalkaline conditions was isolated from hypersaline soda lakes and identifiedas Amphibacillussp. on the basis of 16S rRNA gene sequence analysis.The results showed that Amphibacillussp. strain KSUCr3 was tolerant tovery high Cr(VI) concentration (75 mM) in addition to high tolerance toother heavy metals including Ni 2+ (100 mM), Mo 2+ (75 mM), Co 2+ (5 mM),Mn 2+ (100 mM), Zn 2+ (2 mM), Cu 2+ (2mM) and Pb (75mM). StrainKSUCr3 was shown to be of a high efficiency in detoxifying chromate, asit could rapidly reduce 5 mM of Cr(VI) to a non detectable level over 24 h.In addition, strain KSUCr3 could reduce Cr(VI) efficiently over a widerange of initial Cr(VI) concentrations (1 -10 mM) in alkaline mediumunder aerobic conditions without significant effect on the bacterial growth.Addition of glucose, NaCl and Na 2CO 3 to the culture medium caused adramatic increase in Cr(VI)-reduction by Amphibacillus sp. strainKSUCr3. The maximum chromate removal was exhibited in alkalinemedium containing 1.5% Na 2CO 3, 0.8% glucose, and 1.2% NaCl, atincubation temperature of 40 ºC and shaking of 100 rpm. Under optimumCr (VI) reduction conditions, Cr(VI) reduction rate reached 237 Mh 1which is one of the highest Cr(VI) reduction rate, under alkaline conditionsand high salt concentration, compared to other microorganisms that hasbeen reported so far. Furthermore, the presence of other metals, such asNi 2+ , Co 2+ , Cu 2+ and Mn 2+ slightly stimulated Cr(VI)-reduction ability bythe strain KSUCr3.The isolate,Amphibacillus sp. strain KSUCr3, exhibitedan ability to repeatedly reduce hexavalent chromium without anyamendment of nutrients, suggesting its potential application in continuousbioremediation of Cr(VI). The results also revealed the possible isolationof potent heavy metals resistant bacteria from extreme environment suchas hypersaline soda lakes.OTP003-amylase production by Bacillus species isolated from sweat foodwasteO. Ermithi* 1 , A. Agha 1 , N. Elmarzugi 1,2 , S. Naji 11 Biotechnology Research Center, Microbiology, Tripoli, Libyan ArabJamabiriya2 Alfateh University, Faculty of Pharmacy, Tripoli, Libyan Arab JamabiriyaIndustrial applications of enzymes have been receiving attentionthroughout the world. Amylases are of great importance in biochemicalprocesses, and wide range of application of amylases have used in varioussectors like confectionary, baking, paper, textile, detergent, beverages,baby foods, medicinal and pharmaceutical manufacturing industries whichdrew both researchers and industry excessive attention.It is became a routine work to isolate and produce amylase from differentfungal sources, however, the current work aimed to produce amylaseenzyme from bacterial source (Bacillusspecies). In order to do that, threedifferent formulas has been chosen, the first one is only glucose (starchfree), the second one is mixture of glucose and starch, and the last one isonly starch (with six isolation), at pH 7 (±0.2) and (37°C). The productionactivity has been measured by spectrophotometer in each formula at until8hours time intervals.The results obtained from formula I showed no significant change in thelevel of glucose and this is because of the gene coding of amylase activitywas turned off (enzyme repression) as a result of glucose availability.Formula II showed modest decrease in glucose concentration because thebacteria used the free glucose available rather than breaking up the starchto get glucose. However, in formula III there was an increase in the levelof glucose concentration especially after one hour of incubation as a resultof amylase enzyme activity.Finally, formula III proved the production of -amylase from Bacillusspecies isolated and identified from sweat food waste.1- Kunamneni A, Permaul K, and Singh S (2005) Amylase Production in Solid State Fermentation by theThermophilic FungusThermomyces lanuginosus, Journal of Bioscinence and Bioengineering,100(2): 168-171.2- Akesson M, Hhagander P, and Axelsson J P (2001) -amylase Production in Fed-batch CultivationofBacillus Caldolyticus, Life science Engineering,9: 709.3- Roy A, Moktan B, and Prabir K. Sarar (2007) Characteristics of Bacillus cereus Isolated from legumebasedIndian Fermented Foods,Food Control,18: 1555-15644- Calderon M, loiseau G, and Guyot J P (2003) Fermentation by Lactobacillus Fermentum Ogi E1 ofDifferent Combinations of Carbohydrates Occurring Naturally in Cereals: Consequence on GrowthEnergetic and -amylase ProductionInt. J. Food Microbiol.,80: 161-169.5- Dharani Aiyer P V (2004) Effect of C: N Ratio on Alpha Amylase Production byBacilluslicheniformisSPT 27,African Journal of Biotechnology,V 13(10): 519-522.OTP004Identification of acetate incorporating Arcobacter spp. as potentialmanganese reducers in pelagic redoxclines of the central BalticSea via 16S rRNA based 13 C stable isotope probingC. Berg* 1 , M. Labrenz 1 , S. Beckmann 2 , G. Jost 1 , K. Jürgens 11 Leibniz Institute for Baltic Sea Research (IOW), BiologicalOceanography, Rostock/Warnemünde, Germany2 University of New South Wales (UNSW), School of Biotechnology andBiomolecular Sciences, Sydney, AustraliaPelagic redoxclines in the central Baltic Sea are recognized asenvironments with elevated microbial activities comprising both,heterotrophic and autotrophic prokaryotes, involved in importantbiogeochemical cycles. Aim of our study was to reveal first insights intothe identity and function of heterotrophic bacteria in this habitat which iswell-studied with respect to autotrophic activities. Therefore, pelagicredoxclines of the Gotland basin were sampled in 2005 and 2009,respectively, and subjected to stimulation experiments with differentorganic substrates and electron acceptors, followed by the identification ofstimulated bacteria using 16S rRNA gene single strand conformationpolymorphism (SSCP) analyses. In addition, RNA stable isotope probing(RNA-SIP) followed by subsequent 16S rRNA based quantitative RT-PCRand fingerprinting served to identify acetate incorporating organisms. In2005, in water from the sulfidic zone, 17.3 Mol Mn 4+ were reduced after48 h and bacteria affiliated with the epsilonproteobacterial Arcobacter sp.dominated the incubation. In 2009, bulk incorporation of 3 H labelledacetate was highest in the oxic-anoxic interface layer and still high in thesulfidic zone. After 72 hours, bacteria affiliated with Arcobacter sp.incorporated the 13 C-labeled acetate in the oxic-anoxic interface layer andthe sulfidic zone while the gammaproteobacterial genera Neptunomonassp. and Colwellia sp. incorporated acetate in the oxic-anoxic interfacelayer only. Together, in both experiments two phylogenetically distinctclusters within the genus Arcobacter sp. were identified related topreviously recovered Arcobacter sp. from manganese-oxide rich shelfBIOspektrum | Tagungsband 2012