Molecular Biology Reports

, Volume 46, Issue 4, pp 4565–4580 | Cite as

Preparation and characterization of resistant starch type III from enzymatically hydrolyzed maize flour

  • Anum Khan
  • Ubaid Ur Rahman
  • Samiya Siddiqui
  • Muhammad Irfan
  • Aamer Ali Shah
  • Malik Badshah
  • Fariha Hasan
  • Samiullah KhanEmail author
Original Article


Polysaccharides including resistant starch are categorized as dietary fiber and are used as an important prebiotic. Similar to soluble fibers, resistant starch also has a number of physiological effects that have been shown to be beneficial for health. Starch hydrolyzing enzymes, most importantly amylases, play essential roles in the production of resistant starch. This study aimed to develop α-amylase-treated maize flour with slow digestibility and unique physicochemical characteristics compared to native maize flour. In the current study, resistant starch type III from maize flour was prepared using α-amylase obtained from indigenously isolated Bacillus licheniformis. The α-amylase gene from B. licheniformis was amplified and cloned into the pET-24(a) vector, expressed in E. coli BL21 (DE3) cells and purified by metal ion affinity chromatography. The purified enzyme enhanced the yield of resistant starch 16-fold in maize flour. Scanning electron microscopy revealed that the granular structure of maize flour was disrupted into a dense network with irregular structure, and X-ray diffractograms confirmed the transformation from an amorphous to a crystalline structure upon α-amylase treatment. Thermogravimetric analysis revealed increased amylose content of α-amylase-treated maize flour. Moreover, α-amylase-treated maize flour resulted in a significant enhancement of the desired properties of maize flour, such as resistant starch content, amylose, milk absorption capacity, and iodine and fatty acid complexing ability, and a reduction in swelling power, water binding, oil absorption capacity, and in vitro digestibility compared to untreated maize flour. Resistant starch type III showed low digestibility and increased complexing ability with iodine and fatty acid and therefore could be a safe and beneficial alternative as a coating material for the delivery of active, sensitive ingredients to the colon.


α-Amylase Bacillus licheniformis Resistant starch Slow digestibility Resistant starch with unique characteristics 



The authors thank the Higher Education Commission of Pakistan and TUBITAK, Turkey for providing financial assistance to carry out this work (Grant No. 9-5(Ph-1-MG-4)/Pak-Turk/R&D/HEC/2017.

Compliance with ethical standards

Conflict of interest

We declare no conflict of interest.

Supplementary material

11033_2019_4913_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (DOCX 14 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Microbiology, Faculty of Biological SciencesQuaid-i-Azam UniversityIslamabadPakistan

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