ABSTRACT
The distribution of endo-beta-1,4-glucanase (EG) components in the digestive system of the wood-feeding termite, Coptotermes formosanus Shiraki, was investigated by zymogram analysis using polyacrylamide gel electrophoresis, followed by N-terminal protein sequencing. EG components similar to glycoside hydrolase family (GHF) 9 members were restricted to the salivary glands, the foregut, and the midgut, whereas components similar to GHF7 members were confined to the hindgut where numerous cellulolytic flagellates were harbored. RT-PCR experiments revealed that five GHF9 EG mRNAs (1348 bp) homologous to other termite EGs were expressed in the salivary glands and the midgut. The crude extract prepared from the midgut as well as that from the hindgut produced glucose from crystalline cellulose. These data suggest that C. formosanus has two independent cellulose-digesting systems: one in the midgut where cellulose digestion is accomplished by endogenous cellulases and the other in the hindgut which makes use of other cellulases possibly from symbiotic flagellates.
Subject(s)
Cellulase/metabolism , Cellulose/metabolism , Isoptera/enzymology , Acrylic Resins , Amino Acid Sequence , Animals , Base Sequence , Cellulase/genetics , Cloning, Molecular , DNA, Complementary , Digestive System/enzymology , Isoptera/genetics , Molecular Sequence Data , Sequence Analysis, Protein , Sequence Homology, Amino AcidABSTRACT
Polysaccharide moiety of the boron-polysaccharide complex (T. Matoh, K. Ishigaki, K. Ohno, J. Azuma [1993] Plant Cell Physiol 34: 639-642) isolated from radish (Raphanus sativus) roots has been shown to be rhamnogalacturonan II by glycosyl-linkage analysis and the presence of diagnostic monosaccharides, including apiose, aceric acid, 2-O-methylfucose, and 3-deoxy-D-manno-2-octulosonic acid. Removal of boron from the complex reduced the molecular weight by one-half without causing a significant increase in the number of reducing end groups, indicating that boron, as boric acid, links two rhamnogalacturonan II chains together to form the boron-polysaccharide complex.