occur is evidence of large variations in the rate and extentof starch digestion in the gastrointestinal tract [3–6].Attenuating the ﬂuctuations in postprandial glycaemiaand insulinaemia is important in the prevention and treat-ment of life-style associated diseases, notably diabetesmellitus and cardiovascular disease, and also has implica-tions for obesity management [7–10].The search for full explanations for the differences inthe rates of digestion of starch-containing foods continuesto be a main focus for research, but inter-alia, the differ-encesaremainlyattributedtoinherentdissimilaritiesinthestructure of starch granules of different botanical speciesandfromstructuralalterationssubsequenttodomesticandcommercial processing of foods .Native starch is packaged in granules that are semi-crystallineandwhichareverylargecomparedwiththesizeof the
-amylase molecule. This contrasts with the vastmajority of intracellular metabolic enzymes where theenzyme molecules are usually huge in size compared withthemetabolitesthattheyactupon.Thestarchgranulesizeseemingly presents a very favourable target for attack byamylase with many potential sites for binding of theenzyme. In spite of this apparent binding advantage, thecomplete breakdown of starch within an intact granule is afairly slow process. Crystalline areas tend to be unfavour-able for enzyme attack and, in addition, the granules maycontain small but variable amounts of proteins and lipidsthat can also hinder starch–amylase interaction. Most ofthe starch consumed by humans will have been cookedand/or subjected to various processes during food pro-duction that disrupts the granules to a greater or lesserextent, but raw starch is consumed in many animal feeds.Processing that disrupts general granule integrity andreduces the degree of crystallinity, increases the suscepti-bility to amylase.In recent years, considerable advances have beenmade in knowledge of the structures of starch and mam-malian
-amylasesandmuchhasalsobeenlearnedaboutthe genes coding for
-amylase and their tissue expres-sion. It seems timely, therefore, to review relevant aspectsof what is known about starch and the enzyme to seewhether certain uncertainties in the starch-amylase storycan now begin to be clariﬁed and also perhaps point upaspects of this subject that still need to be resolved byfurther experimentation.
The genes for
-amylase have been cloned by severalgroups and the article by Meiser and Ting  provides anexcellent review of the subject. Salivary glands and thepancreas express the enzyme in abundance but amylaseexpressioncanalsobefoundinthejejunumandmammarygland. Inhumans andother mammals,twodistinctbutclosely related genes code for amylase. AMY1, the sali-vary gene, produces the enzyme present in saliva andmammaryglandandAMY2producestheenzymesynthes-ised in the pancreas that is secreted into the duodenum indigestive juices . The expression of AMY1 can alsooccur in certain tumour tissues, e.g. lung. In humans bothAMY1 and AMY2 are located on the short arm ofchromosome 1 [14–16]. The coding regions of AMY1and AMY2 are very similar but the 5
untranslated regionof AMY2 is shorter than the equivalent region in AMY1. The genes code for enzymes with MWs of about56 000 and the amino acid sequences of the salivaryand pancreatic enzymes differ by only 3%. Both AMY1and AMY2 are polymorphic and alloenzyme forms ofthe salivary and pancreatic amylases have been detected[12, 13]. The two pancreatic forms are exclusive to thepancreas and are expressed at nearly equal levels inthe gland . The secreted salivary amylase has the
-terminal blocked by a pyroglutamic acid residue that may confer some resistance to proteolysis.The organisation of the amylase gene cluster shownbelow is based on published data . The genes arearranged in a head to tail orientation except for AMY1Bwhich is arranged in the reverse order. AMY1P is a pseu-dogene that lacks the ﬁrst three exons of AMY1 .
The promoter regions upstream of all the amylase genescontain a
-actin pseudogene and this ﬁnding has beeninterpreted to indicate that all the amylase genes arederived from a single ancestral gene . In all exceptAMY2B, the
-actin pseudogene is interrupted by a retro-viral-like sequence and this element probably serves aregulatory role in the expression of the salivary enzymes. Meisler and Ting  suggest that an ancestral mam-malian pancreatic gene gave rise to an ancestral AMY2gene which in turn, after
-actin and retroviral insertion,gave rise to the present AMY1 and AMY2 forms.A remarkable feature is that the copy number of theamylasegenesisvariable[12,19].For theAMY1(salivary)gene, variations in copy number from 2 to 15 have beendetected from the screening of 50 individuals .
-Amylase structures and properties
X-ray crystallography has been used to obtain 3D struc-tures of the
-amylase enzymes from human pancreas and saliva  and from porcine pancreas . Thestructures of these enzymes are all very closely related.Mammalian amylases are composed of three structuraldomains, the largest of which (domain A) contains import-ant active site residues that include two aspartate and one396 P. J. Butterworth
et al. Starch/Sta¨ rke
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