Antistaling properties of enzymes (xylanase, bacterial maltogenic and conventional a-amylases), CMC and vital wheat gluten on corn tortillas were evaluated during storage for up to 21 days. Effect of storage time (0-21 days) and temperature (-40, -20, 3, 10 and 21 oC) on tortilla staling was evaluated with or without additives. Addition of 275-1650 AU of ICS maltogenic amylase effectively reduced amylopectin retrogradation without reducing tortilla yields, but did not improve tortilla flexibility. The combination of 825 AU of ICS amylase (to interfere with intra-granular amylopectin re-crystallization) and 0.25% CMC (to create a more flexible inter-granular matrix than retrograded amylose) produced less stiff, equally flexible and less chewy tortillas than 0.5% CMC. Corn tortilla staling followed the basic laws that control aging in starch-based semi-crystalline systems such as starch gels, bread and other baked products. Amylopectin re-crystallization was the driving force behind the staling of corn tortillas. Increasing levels of re-crystallized amylopectin measured by DSC correlated significantly with increased tortilla stiffness and reduction in tortilla rollability, pliability and rupture distance during storage. Re-crystallization of amylopectin in fresh tortillas was not detected. It increased rapidly during the first 24 hr reaching a plateau after 7 days storage. The level of amylopectin re-crystallization on tortillas showed a bell-shaped trend along the evaluated storage temperature range with a maximum around 7 oC. However, a negative linear relationship of peak pasting viscosity with storage temperature of tortilla extracts without additives after 21 days suggests other compounds besides amylopectin affect tortilla staling. Thus, interfering with amylopectin re-crystallization is not the only way to retard staling. Further research is required to optimize the addition of maltogenic amylases in continuous processing lines that use fresh masa instead of nixtamalized corn flour, to determine how these amylases interfere with amylopectin re-crystallization and to elucidate if amylose retrogradation continues during storage and plays a role in tortilla staling.

Master of Science / Department of Grain Science and Industry / Yong Cheng Shi / Maltogenic amylases are widely being used as an antistaling agent in baking industry. However, their action on starch in granular, swelled and dispersed forms, important components formed during bread baking, is largely unknown. Actions of two maltogenic amylases- A and -B on waxy maize starch (WMS) (100% amylopectin) and normal wheat starch (NWS) (~25% amylose) were studied and compared. For any given starch type, starch form, and hydrolysis time, maltogenic amylase-B hydrolyzed both starches more than maltogenic amylase-A as seen through sugar profile analysis indicating its higher degree of multiple attack action (DMA). Their action on non reducing ends blocked compound, p nitrophenol maltoheptaoside, confirmed their endo action. Maltogenic amylase-B showed a higher endo to total enzyme activity ratio than maltogenic amylase-A at any given enzyme weight. Greater MW reduction of dispersed starches by maltogenic amylase-B indicates its higher level of inner chain attack (LICA). Interestingly, MW distributions profiles of swelled starch hydrolysates did not show significant differences irrespective of swelling temperatures. Both enzymes showed differences in oligosaccharides compositions in dispersed and swelled starches’ reaction mixtures with sugars of degree of polymerization (DP) > 2 being degraded to glucose and maltose during later stages. For granular starches, enzymes followed a random pattern of formation and degradation of sugars with DP >2. MW distributions of hydrolyzed granular starches did not show significant shift until at the end of 24h when a low MW peak was observed. Morphological study of granular starches showed that maltogenic amylase-A mainly caused pinholes on WMS while maltogenic amylase-B caused surface corrosion with fewer pinholes. For NWS, both enzymes degraded A granules with deep cavities formation during later stages. A decrease in crystallinity of granular starches means that enzymes were able to hydrolyze both amorphous and crystalline regions. These results indicate that maltogenic amylase-B with a high LICA and high DMA possesses a better starch binding domain which can decrease the starch MW without affecting bread resilience. Strucuture of maltogenic amylase-A modified amylopectin (AP) in relation to its retrogradation was also studied. AP retrogradation was completely inhibited at % DH ≥ 20. MW and chain length distributions of debranched residual AP indicated with increase in % DH, a high proportion of unit chains with DP ≤ 9 and low proportion of unit chains with DP ≥ 17 were formed. Higher proportion of short outer AP chains which cannot participate in double helices formation supports the decrease and eventually complete inhibition of retrogradation. Thus, maltogenic amylase-A can play a very powerful role in inhibiting starch retrogradation even at limited DH (%).