New endo-beta-1,4-glucanases from the parabasalian symbionts, Pseudotrichonympha grassii and Holomastigotoides mirabile of Coptotermes termites.
Abstract. An endo-beta-1,4-glucanase (EG) was purified from the hindgut of an Australian mound-building termite, Coptotermes lacteus. The hindgut extract had a peak separate from those for extracts obtained from the salivary glands and the midgut based on sephacryl S-200 gel chromatography, and also demonstrated an origin different from the endogenous EGs of the termite itself. The recovery was further purified by SDS-PAGE, and its N-terminal amino acid sequence analyzed. This showed high homology to EGs from glycoside hydrolase family (GHF) 7. PCR-based cloning methods were applied to the hindgut contents of C. lacteus and individual protozoan symbionts from C formosanus. cDNAs encoding putative EGs homologous to GHF7 members were then identified. The functionality of one of the putative proteins was confirmed by its expression in Escherichia coli.
Watanabe
H
H
National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8634, Japan. hinabe@affrc.go.jp
Nakashima
K
K
Saito
H
H
Slaytor
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002480:Cellulase / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D056890:Eukaryota / Q000201:enzymology*; D020049:Isoptera / Q000502:physiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D013559:Symbiosis / Q000502:physiology
D018076:DNA, Complementary; D002480:Cellulase