Proteins associated with the surface of wheat starch granules purified by centrifuging through caesium chloride

https://doi.org/10.1016/S0733-5210(09)80157-4Get rights and content

Purified starch was isolated by centrifuging crude starch washed from macerated whole wheat, or an aqueous flour-water suspension, through 80% (w/v) caesium chloride. Granule surface proteins were extracted with 2% sodium dodecylsulphate (SDS) at 20 °C and integral proteins were recovered by extracting with SDS at 50 °C. Integral proteins were qualitatively and quantitatively constant in the starches examined, and provided a convenient internal standard for electrophoresis when total (surface+integral) proteins were extracted together at 50 °C rather than consecutively. In typical starch samples total nitrogen content (35-40 mg N/100 g) was comprised of 3-10 mg N in surface proteins, 13-15 mg N in integral proteins and the 14-20 mg N in lysophospholipids. Electrophoresis on SDS-polyacrylamide gradient (10–20 %) gels showed that purification removed most surface proteins other than the 15 kDa protein friabilin, associated with soft endosperm texture, which was not readily desorbed by the caesium chloride. In starches from soft wheats the small B-granules had more friabilin than the large A-granules, roughly in proportion to their surface area. The method is suitable for screening small samples of wheat for friabilin on its starch and (by association) soft or hard endosperm texture.

References (24)

  • J.B. South et al.

    Cereal Sci.

    (1990)
  • W.R. Morrison et al.

    Cereal Sci.

    (1986)
  • W.R. Morrison

    Anal. Biochem.

    (1964)
  • W.R. Morrison

    J. Cereal Sci.

    (1988)
  • M. Shure et al.

    Cell

    (1983)
  • S.H. Imam

    J. Cereal Sci.

    (1989)
  • P. Greenwell et al.

    Cereal Chem.

    (1986)
  • J.D. Schofield et al.
  • C.W. Wrigley et al.
  • P. Greenwell et al.
  • M. Seguchi et al.

    Cereal Chem,

    (1989)
  • W.R. Morrison et al.

    Cereal Sci.

    (1987)
  • Cited by (55)

    • Making All Medications Gluten Free

      2018, Journal of Pharmaceutical Sciences
      Citation Excerpt :

      Although much of the gluten from the wheat flour separates out during the production of starch as water-insoluble rubbery mass along with certain nongluten proteins, lipids, and carbohydrates, all of the gluten is not removed. Sulaiman and Morrison45 reported that the protein content of starch, which includes gluten, may range from 0.4 to 0.6 percent. The protein content of starch may also vary depending on the manufacturing method and the degree of washing.

    • Viscoelastic properties of tablets from Osborne solubility fraction, pentosans, flour and bread using relaxation tests

      2016, Journal of Cereal Science
      Citation Excerpt :

      MacRitchie (1978, 1985) found that more insoluble HMW fraction (glutenins and residue proteins) was a very critical factor that appeared to account for most of the quality differences between flours of good and poor quality. The protein in the residue of this study more likely contains some of the insoluble glutenin fraction since purified starch contains less than or about 0.4–0.6% protein (Sulaiman and Morrison, 1990). The albumin fraction also presented good viscoelasticity performance.

    • From native malt to pure starch - Development and characterization of a purification procedure for modified starch

      2016, Food Hydrocolloids
      Citation Excerpt :

      Therefore whole kernels were degermed, cracked and steeped in water. The separation of starch from the less dense substances such as cellular material, storage proteins and starch granule surface proteins was achieved by centrifuging through 80% (w/v) cesium chloride solution (South & Morrison, 1990; Sulaiman & Morrison, 1990). Based on the described methods, further purification procedures were developed for different types of cereals (Bohacenko, Chmelik, & Psota, 2006; Ibañez et al., 2007; Peng, Gao, Abdel-Aal, Hucl, & Chibbar, 1999; Pérez, Haros, & Suarez, 2001; Tester, Yousuf, Kettlitz, & Roper, 2007; Wankhede et al., 1990).

    • Use of advanced recombinant lines to study the impact and potential of mutations affecting starch synthesis in barley

      2014, Journal of Cereal Science
      Citation Excerpt :

      The lys5 low donor bulk was generated by pooling equal quantities of grain from three low donor lines; similar low donor bulks were generated for wax from five low donor lines and for isa1 and sex6 from two low donor lines each. Methods were based on those of South and Morrison (1990) and Sulaiman and Morrison (1990). Samples of grain (25 g) were milled to flour in a Perten Laboratory Mill (LM3100, www.perten.com), mixed with 500 ml water at 4 °C for 1 h then filtered through two layers of muslin and one layer of Miracloth.

    View all citing articles on Scopus
    View full text