Comparing the properties of Escherichia coli branching enzyme and maize branching enzyme.
Escherichia coli glycogen branching enzyme (GBE) and maize starch branching enzymes I (SBEI) and II (SBEII) were expressed in E. coli and purified. E. coli GBE branched amylose at a higher rate than did SBEII, but branched amylose at a lower rate than did SBEI. Similar to SBEI, GBE branched amylopectin at a lower rate than did SBEII. High-performance anion-exchange chromatography analysis of the branched products produced by BE revealed the minimum chain length (cl) required for branching. While GBE and SBEII showed the same minimum cl [degree of polymerization (dp) 12] required for branching, SBEI had a slightly higher minimum cl (dp 16) requirement for branching. The major differences between GBE and SBE are their specificities in terms of the size of chains transferred. In comparison with SBE, GBE had a much narrower size range of chains transferred and transferred mainly shorter chains. While SBEI and SBEII produced a large number of chains ranging from dp 6 to over dp 30, GBE predominantly transferred chains ranging from dp 5 to 16 and produced only a very small number of long chains with dp greater than 20. Although it has been reported that SBEI and SBEII preferentially transfer longer and shorter chains, respectively (1), this study further defines the differences between SBEI and SBEII in the size of chains transferred. SBEI predominantly transfers longer chains with dp greater than 10, while producing few shorter chains with dp 3 to 5. In contrast, SBEII preferentially transfers smaller chains with dp 3 to 9, with the most abundant chains being dp 6 and 7. The significance of minimum chain-length requirement by SBE is discussed in setting the invariant size of amylopectin cluster size (9 nm).
Guan
H
H
ExSeed Genetics, Agronomy Department, Iowa State University, Ames 50011, USA. hpguan@iastate.edu
Li
P
P
Imparl-Radosevich
J
J
Preiss
J
J
Keeling
P
P
D015061:1,4-alpha-Glucan Branching Enzyme / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002852:Chromatography, Ion Exchange / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology*; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003313:Zea mays / Q000201:enzymology* / Q000235:genetics
D007527:Isoenzymes; D015061:1,4-alpha-Glucan Branching Enzyme