Starch Branching Enzyme cDNA from Solanum tuberosum

Poulsen, Peter Noe; Kreiberg, Jette D.

doi:10.1104/pp.102.3.1053

Cited by 43 publications

(20 citation statements)

References 6 publications

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“…SSIII primers, 5¢-CACAGGAGGTGTCTGGA AACC-3¢ and 5¢-TGGAACTTGTGAAGGT-GAGGC-3¢ were based on the starch synthase III mRNA sequence (Marshall et al, 1996;X95759). SBEI primers, 5¢-CCGAGCCCCACGAATC-TAT-3¢ and 5¢-GGCTCAGAGCTGCTCATGC-3¢ were based on the starch branching I enzyme mRNA sequence (Poulsen & Kreiberg, 1993;X69805). GBSSI primers, 5¢-GAGCTTCT GGCAGTGAACCC-3¢ and 5¢-GGCAAGTGG AGCGATTTCTTC-3¢ were based on the granule-bound starch synthase I gene sequence (van der Leij et al, 1991;X58453).…”

Section: Immunological Detection Of Dextrans In Tuber Juices and Gelamentioning

confidence: 99%

Production of dextran in transgenic potato plants

et al. 2005

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The production of dextran in potato tubers and its effect on starch biosynthesis were investigated. The mature dextransucrase (DsrS) gene from Leuconostoc mesenteroides was fused to the chloroplastic ferredoxin signal peptide (FD) enabling amyloplast entry, which was driven by the highly tuber-expressed patatin promoter. After transformation of two potato genotypes (cv. Kardal and the amylose-free (amf) mutant), dextrans were detected by enzyme-linked immunosorbent assay (ELISA) in tuber juices of Kardal and amf transformants. The dextran concentration appeared two times higher in the Kardal (about 1.7 mg/g FW) than in the amf transformants. No dextran was detected by ELISA inside the starch granule. Interestingly, starch granule morphology was affected, which might be explained by the accumulation of dextran in tuber juices. In spite of that, no significant changes of the physicochemical properties of the starches were detected. Furthermore, we have observed no clear changes in chain length distributions, despite the known high acceptor efficiency of DSRS.

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Section: Immunological Detection Of Dextrans In Tuber Juices and Gelamentioning

confidence: 99%

Production of dextran in transgenic potato plants

et al. 2005

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“…TCA CAT CAC CAG AAG GAA TAC-3') and downstream P22 primer (5'-CGA TCA ATC ATC TGA CTA GTC-3') were constructed on the basis of comparison of the two previously published cDNA clones, BE7 (KolJmann, 1992) and pSBE8 (Poulsen and Kreiberg, 1993). For amplification of the fulllength clone, the 5' primer (5'-GTT TTG ACC GAT GAC AAT TC-3') and 3' primer (5'-CTA GTC CTC ACC CCA GAC-3') were constructed based on the same comparison as for the P21 and P22 primers.…”

Section: Construction Of Primers the Upstream P21 Primer (5'-mentioning

confidence: 99%

“…To clarify the open reading frame of the 3' end of the sbel gene (KoBmann, 1992;Poulsen and Kreiberg, 1993), a cDNA sequence (sbeI7) coding for the entire mature enzyme was amplified from tuber mRNA of S. tuberosum (cv. Dianella).…”

Section: Identification Of Sbel Allelesmentioning

confidence: 99%

“…Recently, a second isoform of starch-branching enzyme (SBE-11) was detected in preparations of tuber starch from S. tuberosum (Larsson et al, 1996). Further, two full-length cDNA clones of potato SBE-I have been reported, which differ considerably in their 3' translated regions and encode mature enzymes of approximately 95 kDa and 91 kDa (KoBmann, 1992;Poulsen and Kreiberg, 1993). The aim of this work was, firstly, to identify and characterize the active low-molecular-mass form of branching enzyme from S. tuberosum of approximately 80 kDa that was reported earlier (Borovsky et al, 1975;Vos-Scheperkeuter et al, 1989;Blennow and Johansson, 1991) and, secondly, to study the heterogeneity of SBE-I alloforms and the polymorphism of sbeI alleles in S. tuberosum.…”

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The Multiple Forms of Starch‐Branching Enzyme I in Solanum Tuberosum

Khoshnoodi

Blennow

et al. 1996

European Journal of Biochemistry

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Western blot analysis showed the presence of three forms of starch-branching enzyme (SBE), with apparent molecular masses of 103, 97 and 80 kDa, in extracts of leaves and stored tubers of Solanum tuberosurn. The 80-kDa form was absent in extracts of fresh tuber. Active 80-kDa enzyme was partially purified from stored tubers and sequence analysis showed that it, similar to the two larger enzyme forms, was an SBE-I isoform. Limited proteolysis of isolated 103-kDa SBE-I under native conditions removed approximately 200 amino acid residues from the carboxy terminus. A stable intermediate with an apparent molecular mass of approximately 80 kDa was formed. Since the 80-kDa form displayed full enzymatic activity and its circular-dichroism spectrum did not differ significantly from that of the 103-kDa enzyme, the carboxy-terminal portion of the enzyme was suggested to have an extended, unordered structure and therefore to be easily accessible to proteolysis. A cDNA sequence encoding a mature SBE-I was amplified from tuber mRNA of S. tuberosum by means of PCR. The 3' end of this sequence differed significantly from that of previously published data. PCR amplification and DNA sequencing of the 3' ends of the sbel gene showed that four sbel alleles exist in the cultivar studied. Two of these four alleles, sbela and sbelb, had slightly longer 3' ends compared with the other two, sbelc and sbeld. The difference between the two groups of alleles was due to a partial deletion in sbelc and sbeld of a segment duplicated in all alleles. All four alleles were expressed in leaf and tuber.Keywords: starch ; branching enzyme ; allele ; Solanum tuberosum ; Solanurn commersonii.Amylose and amylopectin, the two main components of starch in higher plants, are two a-glucan polysaccharides with different structures and molecular masses. Amylose is essentially a linear (a-1,4 linked) helical molecule with a variable molecular mass (=105-106 Da), while amylopectin, the branched component of the starch, has a significantly higher molecular mass (=107-109 Da) and is the major compound of the starch (Manners, 1985).Starch-branching enzyme (SBE), one of the key enzymes in starch biosynthesis, catalyzes hydrolysis of a-l,4-glucosidic bonds with concomitant condensation of a-1,6-linkages in the amylopectin molecule. Biosynthesis of starch in all plants studied is catalyzed by the same type of enzymes , a number of which, such as starch synthases and branching enzymes, exist as different isoforms, as shown for Zea mays (maize; Ozbun et al

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“…Multiple forms of BE have been identified in many plants, including maize endosperm (4), pea seed (5), and rice endosperm (6 -7). The cDNAs encoding the genes for the BEs have been cloned from various sources, such as maize endosperm (8,9), pea seed (10), potato tuber (11,12), and rice endosperm (6,13). The cDNAs encoding mature mBE I and mBE II have been expressed in Escherichia coli using the T7 promoter (14,15), allowing the study of the structure-function relationships of mBEs using site-directed mutagenesis and the construction of chimeric enzymes of mBE I and mBE II.…”

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Construction of Chimeric Enzymes out of Maize Endosperm Branching Enzymes I and II:

Kuriki¹,

Stewart²,

Preiss

1997

Journal of Biological Chemistry

102

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Branching enzyme I and II isoforms from maize endosperm (mBE I and mBE II, respectively) have quite different properties, and to elucidate the domain(s) that determines the differences, chimeric genes consisting of part mBE I and part mBE II were constructed. When expressed under the control of the T7 promoter in Escherichia coli, several of the chimeric enzymes were inactive. The only fully active chimeric enzyme was mBE II-I BspHI, in which the carboxyl-terminal part of mBE II was exchanged for that of mBE I at a BspHI restriction site and was purified to homogeneity and characterized. Another chimeric enzyme, mBE I-II HindIII, in which the amino-terminal end of mBE II was replaced with that of mBE I, had very little activity and was only partially characterized. The purified mBE II-I BspHI exhibited higher activity than wild-type mBE I and mBE II when assayed by the phosphorylase a stimulation assay. mBE II-I BspHI had substrate specificity (preference for amylose rather than amylopectin) and catalytic capacity similar to mBE I, despite the fact that only the carboxyl terminus was from mBE I, suggesting that the carboxyl terminus may be involved in determining substrate specificity and catalytic capacity. In chain transfer experiments, mBE II-I BspHI transferred more short chains (with a degree of polymerization of around 6) in a fashion similar to mBE II. In contrast, mBE I-II HindIII transferred more long chains (with a degree of polymerization of around 11-12), similar to mBE I, suggesting that the amino terminus of mBEs may play a role in the size of oligosaccharide chain transferred. This study challenges the notion that the catalytic centers for branching enzymes are exclusively located in the central portion of the enzyme; it suggests instead that the amino and carboxyl termini may also be involved in determining substrate preference, catalytic capacity, and chain length transfer.

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Starch Branching Enzyme cDNA from Solanum tuberosum

Cited by 43 publications

References 6 publications

Production of dextran in transgenic potato plants

Production of dextran in transgenic potato plants

The Multiple Forms of Starch‐Branching Enzyme I in Solanum Tuberosum

Construction of Chimeric Enzymes out of Maize Endosperm Branching Enzymes I and II:

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