Identification and preliminary characterization of two cDNAs encoding unique carbonic anhydrases from the marine alga Emiliania huxleyi

被引:61
作者
Sotoj, Amelia R. [1 ]
Zheng, Hong [1 ]
Shoemaker, Dorinda [1 ]
Rodriguez, Jason [1 ]
Read, Betsy A. [1 ]
Wahlund, Thomas M. [1 ]
机构
[1] Calif State Univ San Marcos, Dept Biol Sci, San Marcos, CA 92096 USA
关键词
D O I
10.1128/AEM.00237-06
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Marine coccolithophorid algae are thought to play a significant role in carbon cycling due to their ability to incorporate dissolved inorganic carbon (DIC) into both calcite and photosynthetic products. Among cocco-lithophorids, Emiliania huxleyi is the most prolific, forming massive blooms that affect the global environment. In addition to its ecological importance, the elaborate calcite structures (coccoliths) are being investigated for the design of potential materials for science and biotechnological devices. To date, most of the research focus in this organism has involved the partitioning of DIC between calcification and photosynthesis, primarily using measurements of an external versus internal carbonic anhydrase (CA) activity under defined conditions. The actual genes, proteins, and pathways employed in these processes have not been identified and characterized (see the work of Quinn et al. in this issue [P. Quinn, R. M. Bowers, X. Zhang, T. M. Wahlund, M. A. Fanelli, D. Olszova, and B. A. Read, Appl. Environ. Microbiol. 72:5512-5526, 2006]). In this study, the cloning and preliminary characterization of two genetically distinct carbonic anhydrase cDNAs are described. Phylogenetic analysis indicated that these two genes belonged to the gamma (gamma-EhCA2) and delta (delta-EhCA1) classes of carbonic anhydrases. The deduced amino acid sequence of delta-EhCA1 revealed that it encodes a protein of 702 amino acids (aa) (ca. 77.3 kDa), with a transmembrane N-terminal region of 373 aa and an in-frame C-terminal open reading frame of 329 aa that defines the CA region. The gamma-EhCA2 protein was 235 aa in length (ca. 24.9 kDa) and was successfully expressed in Escherichia coli BL21(DE3) and purified as an active recombinant CA. The expression levels of each transcript from quantitative reverse transcription-PCR experiments under bicarbonate limitation and over a 24-h time course suggest that these isozymes perform different functions in E. huxleyi.
引用
收藏
页码:5500 / 5511
页数:12
相关论文
共 52 条
[1]   Kinetic and spectroscopic characterization of the gamma-carbonic anhydrase from the methanoarchaeon Methanosarcina thermophila [J].
Alber, BE ;
Colangelo, CM ;
Dong, J ;
Stålhandske, CMV ;
Baird, TT ;
Tu, CK ;
Fierke, CA ;
Silverman, DN ;
Scott, RA ;
Ferry, JG .
BIOCHEMISTRY, 1999, 38 (40) :13119-13128
[2]   A CARBONIC-ANHYDRASE FROM THE ARCHAEON METHANOSARCINA-THERMOPHILA [J].
ALBER, BE ;
FERRY, JG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (15) :6909-6913
[3]   Aerobic anoxygenic photosynthesis in Roseobacter clade bacteria from diverse marine habitats [J].
Allgaier, M ;
Uphoff, H ;
Felske, A ;
Wagner-Döbler, I .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2003, 69 (09) :5051-5059
[4]   Costs and benefits of calcification in coccolithophorids [J].
Anning, T ;
Nimer, N ;
Merrett, MJ ;
Brownlee, C .
JOURNAL OF MARINE SYSTEMS, 1996, 9 (1-2) :45-56
[5]  
[Anonymous], PHYCOLOGIA
[6]   CzcD is a heavy metal ion transporter involved in regulation of heavy metal resistance in Ralstonia sp strain CH34 [J].
Anton, A ;
Grosse, C ;
Reissmann, J ;
Pribyl, T ;
Nies, DH .
JOURNAL OF BACTERIOLOGY, 1999, 181 (22) :6876-6881
[7]   The diversity and coevolution of Rubisco, plastids, pyrenoids, and chloroplast-based CO2-concentrating mechanisms in algae [J].
Badger, MR ;
Andrews, TJ ;
Whitney, SM ;
Ludwig, M ;
Yellowlees, DC ;
Leggat, W ;
Price, GD .
CANADIAN JOURNAL OF BOTANY-REVUE CANADIENNE DE BOTANIQUE, 1998, 76 (06) :1052-1071
[8]   Chelation of cellular calcium modulates hypoxia-inducible gene expression through activation of hypoxia-inducible factor-1α [J].
Berchner-Pfannschmidt, U ;
Petrat, F ;
Doege, K ;
Trinidad, B ;
Freitag, P ;
Metzen, E ;
de Groot, H ;
Fandrey, J .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (43) :44976-44986
[9]  
BROWNLEE C, 1994, HAPTOPHYTE ALGAE SYS, V51, P133
[10]   Zinc-bicarbonate colimitation of Emiliania huxleyi [J].
Buitenhuis, ET ;
Timmermans, KR ;
de Baar, HJW .
LIMNOLOGY AND OCEANOGRAPHY, 2003, 48 (04) :1575-1582