Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can after the morphology of amylose-free potato starch granules during biosynthesis

F. Nazarian, G.A. Kok-Jacon, J.P. Vincken, Q. Ji, L.C.J.M. Suurs, R.G.F. Visser

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)

Abstract

It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf background. The starches from the various GtfICAT/SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in physico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and ¿soluble¿ GtfICAT can affect starch biosynthesis differently
Original languageEnglish
Pages (from-to)645-656
JournalTransgenic Research
Volume16
Issue number5
DOIs
Publication statusPublished - 2007

Fingerprint

Sucrase
Amylose
potato starch
Solanum tuberosum
starch granules
amylose
active sites
Starch
Catalytic Domain
biosynthesis
starch
Proteins
proteins
granules
potatoes
amyloplasts
amylopectin
Amylopectin
genetic background
Plastids

Keywords

  • nucleotide-sequence
  • acceptor reactions
  • gtf-i
  • expression
  • plants
  • gene
  • glucosyltransferase
  • dextransucrase
  • glucansucrase
  • reduction

Cite this

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title = "Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can after the morphology of amylose-free potato starch granules during biosynthesis",
abstract = "It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf background. The starches from the various GtfICAT/SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in physico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and ¿soluble¿ GtfICAT can affect starch biosynthesis differently",
keywords = "nucleotide-sequence, acceptor reactions, gtf-i, expression, plants, gene, glucosyltransferase, dextransucrase, glucansucrase, reduction",
author = "F. Nazarian and G.A. Kok-Jacon and J.P. Vincken and Q. Ji and L.C.J.M. Suurs and R.G.F. Visser",
year = "2007",
doi = "10.1007/s11248-006-9053-z",
language = "English",
volume = "16",
pages = "645--656",
journal = "Transgenic Research",
issn = "0962-8819",
publisher = "Springer Verlag",
number = "5",

}

Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can after the morphology of amylose-free potato starch granules during biosynthesis. / Nazarian, F.; Kok-Jacon, G.A.; Vincken, J.P.; Ji, Q.; Suurs, L.C.J.M.; Visser, R.G.F.

In: Transgenic Research, Vol. 16, No. 5, 2007, p. 645-656.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can after the morphology of amylose-free potato starch granules during biosynthesis

AU - Nazarian, F.

AU - Kok-Jacon, G.A.

AU - Vincken, J.P.

AU - Ji, Q.

AU - Suurs, L.C.J.M.

AU - Visser, R.G.F.

PY - 2007

Y1 - 2007

N2 - It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf background. The starches from the various GtfICAT/SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in physico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and ¿soluble¿ GtfICAT can affect starch biosynthesis differently

AB - It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf background. The starches from the various GtfICAT/SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in physico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and ¿soluble¿ GtfICAT can affect starch biosynthesis differently

KW - nucleotide-sequence

KW - acceptor reactions

KW - gtf-i

KW - expression

KW - plants

KW - gene

KW - glucosyltransferase

KW - dextransucrase

KW - glucansucrase

KW - reduction

U2 - 10.1007/s11248-006-9053-z

DO - 10.1007/s11248-006-9053-z

M3 - Article

VL - 16

SP - 645

EP - 656

JO - Transgenic Research

JF - Transgenic Research

SN - 0962-8819

IS - 5

ER -