- Email: [email protected]
Genomic organization, expression and localization of S-adenosylhomocysteine hydrolase from amphioxus Branchiostoma belcheri tsingtaunese
S534
Abstracts / Journal of Biotechnology 136S (2008) S527–S540
Acknowledgement This Work was Supported by the 948 Program of China (2006G55(D)).
References Kumar, S., Tamura, K., Jakobsen, B.I., Nei, M., 2001. MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17, 1244–1245. Yeh, F.C., Yang, R.C., Boyle, T.B., 1997. The User-Friendly Shareware for Population Genetic Analysis. Molecular Biology and Biotechnology Center, University of Alberta, Canada.
doi:10.1016/j.jbiotec.2008.07.1253 VI1-P-019 Genomic organization, expression and localization of Sadenosylhomocysteine hydrolase from amphioxus Branchiostoma belcheri tsingtaunese Yuan Wang 1 , Bosheng Zhao 1,∗ , Shicui Zhang 2 , Xiaojuan Qu 1 1
School of Life Sciences, Shandong University of Technology, Zibo 255049, PR China 2 Department of Marine Biology, Ocean University of China, Qingdao 266003, PR China E-mail address: [email protected] (B. Zhao).
An amphioxus cDNA, encoding S-adenosylhomocysteine hydrolase (AmphiSAHH), was identified from the gut cDNA library of Branchiostoma belcheri tsingtaunese. It contains a 1305-bp open reading frame corresponding to a deduced protein of 434 amino acids. A multiple alignment of SAHH from amphioxus with other known SAHH sequences reveals that some conserved amino acid residues are involved in the unique structural domains within SAHHs. Phylogenetic analysis shows that vertebrate and invertebrate SAHH proteins cluster respectively, with AmphiSAHH falling at the base of vertebrate SAHH clade, which suggests that AmphiSAHH is the archetype of the vertebrate SAHH genes. The amphioxus SAHH gene consists of eight exons flanked by seven introns, which increased gradually during evolution. We express the amphioxus SAHH gene in Escherichia coli driven by T7 promoter. The recombinant amphioxus SAHH protein was purified by HisTrap affinity column. Western blot and immunohistochemistry analysis confirmed that amphioxus has a native molecular mass of approximately 48 kDa, and SAHH was strongly expressed in hepatic caecum, gill, spermary and ovary.
not yet known if the nodules have a biogenic or an abiogenic origin. Here we applied the technique of high-resolution scanning electron microscopy, in combination with energy dispersive X-ray spectroscopical (EDX) analysis, to trace the existence of microbial biofilms in Mn-nodules (Want, in press). Two spatially separated assemblies exist, the exolithobiontic- and endolithobiontic colonizations. The exolithobiontic colonization is seen in the micro-canals, which traverse the outer surface layer of the nodules, and are formed by elongated filamentous organisms, which show no signs of mineralization. In contrast, in the center of the nodules three types of endolithobiontic microbial biofilms exist. First, biofilms formed by cone-like microorganisms, second stone/pillar-like microorganisms and finally paving stone-like, hexagonal microorganisms. They are covered by brick-like mineral deposits. By EDX analysis we could show that on the microorganisms the highest relative level of carbon (C) with respect to manganese (Mn) and sodium can be measured. Our data are in perfect agreement with the assumption that the Mn deposits in the nodules are of biogenic origin. In conclusion, the data presented here, especially those obtained from material taken from the center of the Mn-nodule, indicate that the microorganisms arranged in an organic matrix are at least initially involved in Mn deposition. Deducing from the EDX data it can be concluded that also in the microorganism-free regions a considerable amount of C exists, supporting the view that Mn deposition in Mn-nodule is of biogenic origin. Hence, we strongly suggest a biogenic origin of the Mn-nodules. Next, we will approach the question if in the vicinity of the lithobiontic bacteria, a gradient of Mn(II) to Mn(III)/Mn(IV) exists which could be indicative for the presence of exo-enzymes. If this procedure turns out to be feasible and results in a positive answer we will try to cultivate the endolithobiontic microorganisms in order to enrich the trace amounts of soluble Mn(II) from the marine environment, under enzymatic and oxidative precipitation, into this solid insoluble Mn(IV) to a level which is attractive for biotechnological approaches. Acknowledgements This work was supported by grants from the opening foundation of the Key Laboratory of Marine Sedimentology & Environmental Geology, SOA (grant no. MASEG200602), the China International Science and Technology Cooperation Program (grant no. 20071395), the European Commission and the Bundesministerium für Bildung und Forschung Germany [Center of Excellence BIOTECmarin]. References
Keywords: Amphioxus Branchiostoma belcheri; SAdenosylhomocysteine hydrolase; Characterization; Genome analysis; Expression; Western; Immunohistochemistry
Halbach, P., Friedrich, G., Stackelberg, U., 1988. The Manganese Nodule Belt of the Pacific Ocean. Enke, Stuttgart. Wang, X., Schloßmacher, U., Wiens, M., Schröder, H.C., Müller, W.E.G., in press. Biogenic origin of polymetallic nodules from the Clarion-Clipperton Zone in the Eastern Pacific Ocean: Electron microscopic and EDX evidence. Mar. Biotechnol.
doi:10.1016/j.jbiotec.2008.07.1254
doi:10.1016/j.jbiotec.2008.07.1255
VI1-P-020
VI1-P-021
Biotic colonizations of manganese/polymetallic nodules
The phylogenetic implications of cephalopods based on 16S rRNA sequences
Xiaohong Wang ∗ , Yimin Wang, Werner E.G. Müller National Research Center for Geoanalysis, 26 Baiwanzhuang Dajie, CHN-100037 Beijing, China E-mail addresses: [email protected] (X. Wang), [email protected] (W.E.G. Müller). Polymetallic/ferro-manganese nodules (Mn-nodules) provide a rich source for manganese (Halbach et al., 1988; Want, in press). It is
Jingni Hong ∗ , Jia-ying Du, Yong Mao, Jun Wang Department of Oceanography, Xiamen University, Xiamen 361005 China E-mail address: [email protected] (J. Hong). Some comprehensive analysis were conducted based on molecular data such as the sequences of mt DNA and nuclear DNA gene
Abstracts / Journal of Biotechnology 136S (2008) S527–S540
Acknowledgement This Work was Supported by the 948 Program of China (2006G55(D)).
References Kumar, S., Tamura, K., Jakobsen, B.I., Nei, M., 2001. MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17, 1244–1245. Yeh, F.C., Yang, R.C., Boyle, T.B., 1997. The User-Friendly Shareware for Population Genetic Analysis. Molecular Biology and Biotechnology Center, University of Alberta, Canada.
doi:10.1016/j.jbiotec.2008.07.1253 VI1-P-019 Genomic organization, expression and localization of Sadenosylhomocysteine hydrolase from amphioxus Branchiostoma belcheri tsingtaunese Yuan Wang 1 , Bosheng Zhao 1,∗ , Shicui Zhang 2 , Xiaojuan Qu 1 1
School of Life Sciences, Shandong University of Technology, Zibo 255049, PR China 2 Department of Marine Biology, Ocean University of China, Qingdao 266003, PR China E-mail address: [email protected] (B. Zhao).
An amphioxus cDNA, encoding S-adenosylhomocysteine hydrolase (AmphiSAHH), was identified from the gut cDNA library of Branchiostoma belcheri tsingtaunese. It contains a 1305-bp open reading frame corresponding to a deduced protein of 434 amino acids. A multiple alignment of SAHH from amphioxus with other known SAHH sequences reveals that some conserved amino acid residues are involved in the unique structural domains within SAHHs. Phylogenetic analysis shows that vertebrate and invertebrate SAHH proteins cluster respectively, with AmphiSAHH falling at the base of vertebrate SAHH clade, which suggests that AmphiSAHH is the archetype of the vertebrate SAHH genes. The amphioxus SAHH gene consists of eight exons flanked by seven introns, which increased gradually during evolution. We express the amphioxus SAHH gene in Escherichia coli driven by T7 promoter. The recombinant amphioxus SAHH protein was purified by HisTrap affinity column. Western blot and immunohistochemistry analysis confirmed that amphioxus has a native molecular mass of approximately 48 kDa, and SAHH was strongly expressed in hepatic caecum, gill, spermary and ovary.
not yet known if the nodules have a biogenic or an abiogenic origin. Here we applied the technique of high-resolution scanning electron microscopy, in combination with energy dispersive X-ray spectroscopical (EDX) analysis, to trace the existence of microbial biofilms in Mn-nodules (Want, in press). Two spatially separated assemblies exist, the exolithobiontic- and endolithobiontic colonizations. The exolithobiontic colonization is seen in the micro-canals, which traverse the outer surface layer of the nodules, and are formed by elongated filamentous organisms, which show no signs of mineralization. In contrast, in the center of the nodules three types of endolithobiontic microbial biofilms exist. First, biofilms formed by cone-like microorganisms, second stone/pillar-like microorganisms and finally paving stone-like, hexagonal microorganisms. They are covered by brick-like mineral deposits. By EDX analysis we could show that on the microorganisms the highest relative level of carbon (C) with respect to manganese (Mn) and sodium can be measured. Our data are in perfect agreement with the assumption that the Mn deposits in the nodules are of biogenic origin. In conclusion, the data presented here, especially those obtained from material taken from the center of the Mn-nodule, indicate that the microorganisms arranged in an organic matrix are at least initially involved in Mn deposition. Deducing from the EDX data it can be concluded that also in the microorganism-free regions a considerable amount of C exists, supporting the view that Mn deposition in Mn-nodule is of biogenic origin. Hence, we strongly suggest a biogenic origin of the Mn-nodules. Next, we will approach the question if in the vicinity of the lithobiontic bacteria, a gradient of Mn(II) to Mn(III)/Mn(IV) exists which could be indicative for the presence of exo-enzymes. If this procedure turns out to be feasible and results in a positive answer we will try to cultivate the endolithobiontic microorganisms in order to enrich the trace amounts of soluble Mn(II) from the marine environment, under enzymatic and oxidative precipitation, into this solid insoluble Mn(IV) to a level which is attractive for biotechnological approaches. Acknowledgements This work was supported by grants from the opening foundation of the Key Laboratory of Marine Sedimentology & Environmental Geology, SOA (grant no. MASEG200602), the China International Science and Technology Cooperation Program (grant no. 20071395), the European Commission and the Bundesministerium für Bildung und Forschung Germany [Center of Excellence BIOTECmarin]. References
Keywords: Amphioxus Branchiostoma belcheri; SAdenosylhomocysteine hydrolase; Characterization; Genome analysis; Expression; Western; Immunohistochemistry
Halbach, P., Friedrich, G., Stackelberg, U., 1988. The Manganese Nodule Belt of the Pacific Ocean. Enke, Stuttgart. Wang, X., Schloßmacher, U., Wiens, M., Schröder, H.C., Müller, W.E.G., in press. Biogenic origin of polymetallic nodules from the Clarion-Clipperton Zone in the Eastern Pacific Ocean: Electron microscopic and EDX evidence. Mar. Biotechnol.
doi:10.1016/j.jbiotec.2008.07.1254
doi:10.1016/j.jbiotec.2008.07.1255
VI1-P-020
VI1-P-021
Biotic colonizations of manganese/polymetallic nodules
The phylogenetic implications of cephalopods based on 16S rRNA sequences
Xiaohong Wang ∗ , Yimin Wang, Werner E.G. Müller National Research Center for Geoanalysis, 26 Baiwanzhuang Dajie, CHN-100037 Beijing, China E-mail addresses: [email protected] (X. Wang), [email protected] (W.E.G. Müller). Polymetallic/ferro-manganese nodules (Mn-nodules) provide a rich source for manganese (Halbach et al., 1988; Want, in press). It is
Jingni Hong ∗ , Jia-ying Du, Yong Mao, Jun Wang Department of Oceanography, Xiamen University, Xiamen 361005 China E-mail address: [email protected] (J. Hong). Some comprehensive analysis were conducted based on molecular data such as the sequences of mt DNA and nuclear DNA gene