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Starch: Structure, Function, and Biosynthesis Outline
Introduction to starch Starch biosynthesis and genetic modifications Starch analysis Functional properties of food starch Modification of starches Starch digestibility

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Introduction to Starch Starch
ƒ Carbon and energy storage material for plants ƒ A major component of food and feed, providing source of energy and carbon ƒ Used for texturizing and to provide specific functionalities in processed foods ƒ Starting material for ethanol production ƒ Broad applications in the paper and textile industries

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Introduction to Starch Industrial Starch Sources Corn starch
ƒ ƒ ƒ ƒ Common corn Waxy maize (amylose ~ 0%) High-amylose (amylose 55 and 70%) Other mutants

Root/Tuber starch
ƒ Potato ƒ Cassava (Tapioca)

Other Cereals
ƒ Wheat ƒ Rice ƒ Barley

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Introduction to Starch
Partial list of documented starch mutants of maize
Single mutants
‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ Waxy (wx) Amylose-extender (ae) sbe1 sbe2a Sugary-1 (su1) Zpu1 Sugary-2 (su2) Dull-1 (du1) Brittle-1 (bt1) Brittle-2 (bt2) Shrunken-1 (sh1) Shrunken-2 (sh2)

double mutants
‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ae wx sbe1 wx ae su1 ae su2 ae du1 du1 su1 dul su2 du1 wx su1 wx su2 wx su1 su2

Triple/quadruple mutants
‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ae du1 su1 ae du1 su2 ae du1 wx ae su1 su2 ae su1 wx ae su2 wx sbe1 ae wx du1 su1 su2 du1 su1 wx du1 su2 wx su1 su2 wx ae du1 su1 wx

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Introduction to Starch World starch production and utilizations
Starch Production Corn Potato Wheat Tapioca Others 83% 6% 6% 4% 1% Refinery Products Dextrose and maltose High fructose corn syrup Maltodextrin Ethanol Starch Products Paper (ca. 60%) Food (ca. 20%) Textile (ca. 20%)

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Introduction to Starch End-use demand for starch in major markets
Approx. % US Paper and paperboard High-fructose sweetener Other sweeteners Fuel ethanol Food Other industrial
a Corn

EU 22 5 28 2 18 26

Japan 11 27 27 5 21 10

10 31 12 42 3 2

Annual 2001, Corn Refiners association

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Introduction to Starch Starch granules

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Introduction to Starch Starch granules

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Introduction to Starch Starch granules

Normal light

Polarized light

Birefringence indicates that the granules are semi-crystalline

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Introduction to Starch Starch granules

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Introduction to Starch Starch granules

Two populations of wheat starch: large size A granules, and small size B granules

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Introduction to Starch Starch granule size distributions

85nm 10-15 nm in d iam eter 14 Amylose and Amylopectin Amylose ‰ A linear molecule comprising of 1.000 .04nm C olum n of w ater 120 0 1.4 linked and 1.000 nm Overview of Starch Granule C ry sta lline H ard S he ll (120-400 nm thick) S em i-crysta lline S oft S hell H ilum 20-500 nm C rysta lline hard she ll W h o le G ra n ule P o res (100-300 nm in d iam eter) G ranule S u rfac e S u p e rh e lix o f A P T ilte d C lu sters S id e P la n C rysta lline Lam ellae A m orpho us Lam ellae A xis o f AP helices S em i-crysta lline ha rd she ll A P double he lice s A m o rp h o u s c h a n n e ls P itc h of lam ella r he lice s (9 nm ) b locklet b locklet 10-50 nm & 1.12nm T o p V ie w 123.85nm 1.13 1.17nm ~1 nm Am ylopec tin C lu ster AM L ipid L ip id 1nm 6nm T y pe A 3nm T y pe B AM 1. There is a small degree of branching by 1.6 alpha linkages ‰ The smaller of the two polysaccharides making up starch Amylopectin ‰ A highly branched molecule comprising both 1.6 linked alpha-D-glucopyranosyl units ‰ Branches are non-randomly distributed in clusters ‰ The larger of the two polysaccharides making up starch 15 Amylose and Amylopectin Diagrams of amylopectin branches & clusters .5 0 1.5 c lusters B lo ck let (S ize 50 -500 nm depend ing o n bota nica l so urce a nd loc atio n in gra nule ) 9-10n m O ute r rad ius (9 nm ) T o p P la n P ie-shaped Lam ellar M otif Inne r R ad ius (4 nm ) C entra l ho le w ith the sam e dens ity as am orp ho us lam e llae 1.100.07nm S ide V iew 2.4 linked alpha-D-glucopyranosyl units.

chains connecting three clusters B1a chain. 481 18 Amylose and Amylopectin Structural parameters of amylopectin clusters ICL of B3 (53. long intra-cluster chains A chains Yao et al. 136: 3515–3523 . B and C ‰ A chains are not branched ‰ B chains are branched ‰ C chain has the reducing end R ‰ One reducing end per amylopectin molecule Wang et. 2004.2 DP) B2 chain.16 Amylose and Amylopectin Reducing end Open ring Aldehyde oxidized to carboxylic acid 17 Amylose and Amylopectin Different type of chains for amylopectin ‰ Three chain types A. Botany (1998) 49. Plant Physiology.5 DP) wx wx β-limit dextrin B chain stub 1 or 2 DP A chain stub 2 or 3 DP ICL of B1-a (3. short intra-cluster chains B1b chain. al. chains connecting two clusters B3 chain. J. Exp..5 DP) Average number of branches per cluster (ANBPC) ICL of B2 (26.5 DP) ae wx Cluster repeat distance (26.

85 20 Starch Crystallinity Crystalline forms ‰ A: Cereal starch ‰ B: Tuber starch. Int. water molecules. J. Macromol (1998) 23.. iodine. Macromol. (1998) 23. Int. An (a. etc ‰ Vh: hydrated V form Buleon et al. 85 21 Starch Crystallinity Crystalline A structure Crystalline A structure for starch.. Biol. (1988) 201. J. Biol. 365 . high amylose starch ‰ V: amylose crystallized with lipids. Mol. and hydrogen bonding Imberty et al.b) plane projection of the unit showing helix packing. Biol.19 Amylose and Amylopectin Starch-lipid complex Polar (hydrophilic) head outside Nonpolar (hydrophobic) tail inside Buleon et al. J.

Biopolymers (1988) 27. Rengaswami Chandrasekaran 24 Starch Biosynthesis and Genetic Starch Modifications of Maize .b) plane projection of the unit showing nearby helices and the center channel of organized water molecules Imberty et al. An (a.22 Starch Crystallinity Crystalline B structure Crystalline B structure for starch.1205 23 Amylose and Amylopectin Estimation of Crystallinity α Amorphous Crystalline area = 435 α = 435 / 1460 = 30% Total area = 1460 Diagram from Dr.

25 Outline ‰ An introduction from maize kernel to amylopectin clusters ‰ Functional behaviors of enzymes synthesizing starch ‰ Mutant starches of maize. single mutants ‰ Mutant starches of maize. Illinois. Chicago. and an industrial starting material. an energy storage for germination. ‰ Starch forms in leaf chloroplasts during the day ‰ And hydrolyzed and transported to other plant parts at night as simple sugar ‰ Reserve starch.g. e. patents associated with foods ‰ Genetic starch modifications. 1964 . is formed in amyloplasts ‰ We will be discussing starch biosynthesis in maize endosperm 27 From Maize Kernel to Amylopectin Clusters A kernel of yellow dent corn Diagram adapted from Wheat Flours Institute. multiple mutants ‰ Mutant starches of maize. our approaches 26 From Maize Kernel to Amylopectin Clusters Starch formation in plastids ‰ Starch granules are classified as transitory or reserve ‰ Transitory starch granules accumulate for only a short period of time before they are degraded. challenges ‰ Genetic starch modifications. a major component of food and feed.

1997. 32:177-191 30 From Maize Kernel to Amylopectin Clusters Amylopectin branches are non-randomly clustered Thompson. and starch molecular structures 29 From Maize Kernel to Amylopectin Clusters Let’s look at the big picture before “zooming in”…… We are going here! Gallant et al. Carbohydrate Polymers.28 From Maize Kernel to Amylopectin Clusters Comparison of kernels and starch granules of two wx mutants wx ae wx Genetic modifications provide diverse kernel phenotypes. 43: 223-239 . internal granular structures. starch granule sizes. 2000. Carbohydrate Polymer.

min 33 From Maize Kernel to Amylopectin Clusters What can genetic starch modifications do for us? ‰ Improve the starch yield of major crops ‰ Acquire starches with desirable functionalities and high value ‰ Suitable starting materials for chemical and enzymatic modifications ‰ Modified digestibility or degradability as food.I 60 40 20 0 10 12 14 16 18 20 B3 ICL + 2. A chains. feed. min 12 14 16 18 20 RETENTION TIME. as carriers for controlled release .31 From Maize Kernel to Amylopectin Clusters Structural parameters of amylopectin clusters Keywords: amylopectin.5 40 20 B1-a ICL+ 2. branching pattern ICL of B3 (53. 2004. Plant Physiology. clusters. and industrial raw materials ‰ Retarded or enhanced retrogradation after cooking.5 DP) wx wx β-limit dextrin B chain stub 1 or 2 DP A chain stub 2 or 3 DP ICL of B1-a (3. branches.R.5 B1-b ICL+ 2.5 B2 B1-b B1-a A B2 ICL+ 2. DP 60 NORMALIZED D. B chains. e.5 DP) Average number of branches per cluster (ANBPC) ICL of B2 (26. DP 1000 100 50 20 10 3 Debranched β-limit dextrin DEGREE of POLYMERIZATION. 136: 3515–3523 32 From Maize Kernel to Amylopectin Clusters Chain length profiles are used to calculate AP parameters Debranched amylopectin DEGREE of POLYMERIZATION.R. leading to extended shelf life or unique functionalities ‰ Unique granular and nano structures for high-end uses.g.5 DP) ae wx Cluster repeat distance (26.2 DP) Yao et al. DP ae wx wx RELATIVE MOLAR AMOUNT DEGREE of POLYMERIZATION. DP RELATIVE MOLAR AMOUNT 5 4 3 2 1 0 10 1000 100 50 20 10 3 15 1000 100 50 B3 B2 20 10 B1-b B1-a 3 A Average CL 10 A chain amount Cluster amount 5 B1-a ICL+ 2.5 0 10 12 14 16 18 20 RETENTION TIME.I 1000 100 50 B3 20 10 3 NORMALIZED D.5 0 10 12 14 16 18 20 DEGREE of POLYMERIZATION.

34 From Maize Kernel to Amylopectin Clusters Successful genetic starch modification is based on the knowledge of starch biosynthesis ‰ Where are starch granules synthesized? ‰ What enzymes are involved in starch biosynthesis? ‰ What are the functional behaviors of these enzymes? ‰ Can we tailor starch structure for desirable functions? 35 Enzymes Synthesizing Starch Starch biosynthesis in a cell of maize endosperm Starch debranching enzyme Starch branching enzyme starch granules amylopectin + amylose ADP AMYLOPLAST D-enzyme? Soluble starch synthase Granule-bond starch synthase Glucosyl primer (UDP-Glc:protein transglucosylase?) ADP-Glc transporter ADP-Glc ADP-Glc ATP Phosphoglucomutase ATP PPi ADP-Glc pyrophosphorylase UTP UDP-Glc pyrophosphorylase ADP Glc-1-P + PPi Fructose UDP-Glc Sucrose synthase Sucrose UDP CYTOSOL 36 Enzymes Synthesizing Starch ADP-glucose pyrophosphorylase (AGPase) ATP PPi O P AGPase ADP-glucose ADP .

37 Enzymes Synthesizing Starch Starch synthase (SS) Isoforms identified: GBSSI. SSIIa. SSI. SBEIIa. & SBEIIb O O O O O O Reaction I O O SBE O O O O O O O + O O O O O O O O Reaction II O O SBE O O O O O O + O O O 39 Enzymes Synthesizing Starch Starch debranching enzyme (DBE) Isoforms identified: Isoamylase-like (SU1) & Pullulanase-like (ZPU1) O O O O O O DBE O O O O O . SSIIb. & SSIII + ADP O O O O SS ADP O O O O O ‰ Soluble starch synthase (SSS) responsible for amylopectin synthesis ‰ Granule-bond starch synthase (GBSS) responsible for amylose synthesis 38 Enzymes Synthesizing Starch Starch branching enzyme (SBE) Isoforms identified: SBEI.

40 Enzymes Synthesizing Starch Specific substrates may detect enzyme specificity Plant tissue Protein extraction Native PAGE Electroblotting Incubation Iodine staining Photo recording SBEIa SS-4 SBEIIa SBEIIb DBE SS-5 SS-6 ae wx sbe1a ae wx wx sbe1a wx ae wx × 1/2 Glucan Glucan dispersion Glucan gel Potato starch sbe1a ae wx × 1/2 Waxy maize starch sbe1a ae wx × 1/2 ae wx sbe1a ae wx wx sbe1a wx ae wx × 1/2 Liver glycogen wx sbe1a wx ae wx sbe1a ae wx wx × 1/2 sbe1a wx × 1/2 ae wx × 1/2 sbe1a ae wx × 1/2 SS-1 SS-2 SS-3 41 Enzymes Synthesizing Starch Starch branching enzymes change gel from blue to purple or yellow sbe1a ae wx × 1/2 sbe1a ae wx × 1/2 wx sbe1a wx ae wx sbe1a ae wx wx × 1/2 sbe1a wx × 1/2 ae wx × 1/2 sbe1a ae wx × 1/2 SS-1 SS-2 SS-3 SBEIa SS-4 SBEIIa SBEIIb DBE SS-5 SS-6 ae wx sbe1a ae wx wx sbe1a wx ae wx sbe1a ae wx wx sbe1a wx ae wx × 1/2 Potato starch ‰ ‰ ‰ ae wx × 1/2 Waxy maize starch Liver glycogen All SBEs may branch both amylose & amylopectin SBEI preferentially branches amylose SBEIIa & SBEIIb preferentially branches amylopectin 42 Enzymes Synthesizing Starch Starch debranching enzymes change gel from purple to blue sbe1a ae wx × 1/2 sbe1a ae wx × 1/2 wx sbe1a wx ae wx sbe1a ae wx wx × 1/2 sbe1a wx × 1/2 ae wx × 1/2 sbe1a ae wx × 1/2 SS-1 SS-2 SS-3 SBEIa SS-4 SBEIIa SBEIIb DBE SS-5 SS-6 ae wx sbe1a ae wx wx sbe1a wx Potato starch ‰ ‰ ae wx × 1/2 Waxy maize starch ae wx sbe1a ae wx wx sbe1a wx ae wx × 1/2 Liver glycogen Shown here is a pullulanase-like DBE (ZPU1) An isoamylase-like DBE (SU1) is not shown .

rice. e. Single Mutants Waxy (wx) ‰ Deficiency of granule-bond starch synthase ‰ Identified in maize.43 Enzymes Synthesizing Starch Starch synthases change gel from yellow to dark blue sbe1a ae wx × 1/2 sbe1a ae wx × 1/2 wx sbe1a wx ae wx sbe1a ae wx wx × 1/2 sbe1a wx × 1/2 ae wx × 1/2 sbe1a ae wx × 1/2 SS-1 SS-2 SS-3 SBEIa SS-4 SBEIIa SBEIIb DBE SS-5 SS-6 ae wx sbe1a ae wx wx sbe1a wx Potato starch ‰ ‰ ae wx × 1/2 Waxy maize starch ae wx sbe1a ae wx wx sbe1a wx ae wx × 1/2 Liver glycogen At least 5 bands are shown here.g. multiple mutants containing wx has NO amylose ‰ Dosage effect shown for Wx wx wx ‰ Broad applications in the food and non-food industries . wheat. & potato ‰ Kernels of wx are full ‰ Starch and dry weight are equal to normal ‰ Mutant wx is epistatic to other known mutants. sorghum. barley. with different activities We will assign these bands to known SS isoforms 44 Mutant Starches from Maize A list of documented starch mutants of maize Single mutants ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ Waxy (wx) Amylose-extender (ae) sbe1 sbe2a Sugary-1 (su1) Zpu1 Sugary-2 (su2) Dull-1 (du1) Brittle-1 (bt1) Brittle-2 (bt2) Shrunken-1 (sh1) Shrunken-2 (sh2) double mutants ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ae wx sbe1 wx ae su1 ae su2 ae du1 du1 su1 dul su2 du1 wx su1 wx su2 wx su1 su2 Triple/quadruple mutants ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ae du1 su1 ae du1 su2 ae du1 wx ae su1 su2 ae su1 wx ae su2 wx sbe1 ae wx du1 su1 su2 du1 su1 wx du1 su2 wx su1 su2 wx ae du1 su1 wx 45 Mutant Starches.

Single Mutants Sugary-1 (su1) ‰ Deficiency of starch debranching enzyme SU1 ‰ Identified in maize. Single Mutants Amylose-Extender (ae) ‰ Deficiency of starch branching enzyme IIb (SBEIIb) for maize ‰ Identified in maize. Amylopectin has increased proportion of short chains ‰ Thermal properties by DSC are related to su2 dosage . Chromatographic analysis shows lower amylose value and the presence of intermediate materials ‰ Granules are smaller than normal. and some may be nonbirefringent ‰ B-type x-ray pattern is shown for ae starch ‰ Broad applications 47 Mutant Starches. peas. & barley ‰ Kernels of ae have a smaller size than normal ‰ Dosage effect exists for ae gene ‰ Amylose concentration usually ranges 50-75% by blue value tests. & rice ‰ Standard sweet corn is su1 homozygous ‰ Kernels of su1 accumulates phytoglycogen >25% of kernel dry weight. Single Mutants Sugary-2 (su2) ‰ Possible deficiency of a starch synthase SSIIa ‰ Identified in maize. rice. Small starch granules may be isolated with amylose content possibly higher than normal ‰ Kernel development shows a pattern of: Starch granules formed → granules partially degraded and replaced by phytoglycogen → granules mostly degraded and replaced by phytoglycogen ‰ Kernels have intermediate particles ranging from amylopectin to phytoglycogen 48 Mutant Starches.46 Mutant Starches. possible in sorghum & rice ‰ Kernel dry weight is often reduced ‰ Starch granules have extensive internal fractures ‰ Amylose content is 10-15% higher than normal. sorghum.

Multiple Mutants Amylose-Extender Sugary-1 (ae su1) ‰ Deficiency of both SBEIIb and SU1 ‰ Kernels not as full as ae. but chromatographic separation indicates solely amylopectin ‰ Both ae and wx functions independently ‰ Amylopectin has increased proportion of long chains ‰ Dosage effect exists for both ae and wx genes 51 Mutant Starches. Weight and starch concentration are lower than normal ‰ Increased doses of ae results in reduced phytoglycogen ‰ Blue value test indicates 51-60% amylose content ‰ In su1. Single Mutants Dull-1 (du1) ‰ Deficiency of a starch synthase SSIII ‰ Kernel ranging from full size to semi-collapsed ‰ Average granule size smaller than normal ‰ Amylose content 5-10% higher than normal.49 Mutant Starches. but fuller than su1. In ae su1. Multiple Mutants Amylose-Extender Waxy (ae wx) ‰ Deficiency of both SBEIIb and GBSSI ‰ Reduced in size. the initially formed starch granules are broken down and may be utilized to produce phytoglycogen. and starch content (~50%) ‰ Blue value test indicates 15-26% amylose. dry weight. Amylopectin has increased proportion of short chains ‰ Mutant du1 in sweet corn background may accumulate phytoglycogen 50 Mutant Starches. ae interferes with excessive branching and starch granules are formed along with small amount of phytoglycogen .

min wx du1 wx ae wx du1 ae wx 12 14 16 18 20 RETENTION TIME. Sugar contents are several fold higher. 70 NORMALIZED D. and somewhat intermediate between wx and du1 wx 54 Mutant Starches. and starch concentration is lower than in either normal. Multiple Mutants Amylose-Extender Dull-1 Waxy (ae du1 wx) ‰ Deficiency of SBEIIb. SSIII. wx. Multiple Mutants Amylopectin of mutants containing ae. so du1 wx has 100% of amylopectin ‰ Amylopectin has increased portion of shorter chains 53 Mutant Starches. Sweetness is between standard sweet corn (su1) and sh2 mutation.52 Mutant Starches.R. slightly less than normal ‰ Sugar concentrations are higher. 60 50 40 30 20 10 0 10 1000 100 50 20 10 3 60 50 40 30 20 10 0 10 12 14 16 18 20 RETENTION TIME.I. du1. ‰ Amylopectin branching is different from either ae wx or du1 wx. and GBSSI ‰ Starch concentration is low compared with the component single and double mutants.I. Multiple Mutants Dull-1 Waxy (du1 wx) ‰ Deficiency of both SSIII and GBSSI ‰ Kernels weight similar to du1 and wx. or wx ‰ Gene wx is epistatic to du1. min . and du1 Chain length profile of amylopectin Chain length profile of β-limit dextrin DEGREE of POLYMERIZATION DEGREE of POLYMERIZATION 1000 100 50 20 10 3 NORMALIZED D.R.

Foodstuffs containing starch of an amylose extender sugary-2 genotype ‰ 1988. Patents A list of mutant starch patents in last 20 years ‰ 2004. Bread containing wx su2 genotype starch as an anti-stalent 57 Mutant Starches. ‰ ‰ ‰ ‰ . Starch of the wx sh1 genotype and products produced therefrom ‰ 1986. Foodstuffs containing starch of a waxy shrunken-2 genotype ‰ 1989. Food stuffs containing starch of a dull sugary-2 genotype ‰ 1988. Food stuffs containing starch of a dull waxy genotype ‰ 1988. gelling. The samples were visually inspected for opacity.474) Starch A B waxy 2 doses ‰ Opacity 3 2 <1 3 Gelling 3 2 <1 3 Syneresis (Surface) 3 2 <1 3 Syneresis (Pressed) 3 2 <1 3 Overall stability 3 2 <1 3 Freeze-thaw stability: starch pastes were placed at -19C for 16 hours and then thawed for 8 hours at room temperature. ae du1 batter starch for deep fat fried food ‰ 1989.828.55 Mutant Starches. Method of grain production for heterozygous waxy sugary-2 maize ‰ 1996. the Potential Potential of mutant combinations Single mutants ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ Waxy (wx) Amylose-extender (ae) sbe1 sbe2a Sugary-1 (su1) Zpu1 Sugary-2 (su2) Dull-1 (du1) Brittle-1 (bt1) Brittle-2 (bt2) Shrunken-1 (sh1) Shrunken-2 (sh2) double mutants ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ae wx sbe1 wx ae su1 ae su2 ae du1 du1 su1 dul su2 du1 wx su1 wx su2 wx su1 su2 Triple/quadruple mutants ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ae du1 su1 ae du1 su2 ae du1 wx ae su1 su2 ae su1 wx ae su2 wx sbe1 ae wx du1 su1 su2 du1 su1 wx du1 su2 wx su1 su2 wx ae du1 su1 wx 56 Mutant Starches. Starch of wx fl1 genotype and products produced therefrom ‰ 1988. Patents An example: Double Mutant Homozygous wx Heterozygous su2 Freeze-thaw stability of different starches (US patent # 6. Starch of the duh genotype and products produced therefrom ‰ 1988. Foodstuffs containing a waxy waxy amylose extender starch ‰ 1992. Food stuffs containing starch of an amylose extender dull genotype ‰ 1988. and syneresis (both on the surface and when pressed) Sample A: blend of approximately 10% waxy maize and 90% waxy maize with two doses of su2 gene Sample B: blend of approximately 20% waxy maize and 80% waxy maize with two doses of su2 gene 2 doses: pure waxy maize hybrid with two doses of su2 gene Each number represents the number of freeze-thaw cycles the starch paste remained acceptable (stable).

79: 757–762 .1 0. DP FACE by MKI by SKS Yao et al.02 0 0 5 10 15 20 25 30 35 40 45 50 55 60 DEGREE OF POLYM ERIZATION.08 0.06 0. The Challenges Our strategy to generate novel starches Starch functionalities desirable by industries Starch structure required for desirable functionalities Starch materials for chemical & enzymatic modifications Methodologies connecting plant genetics with carbohydrate chemistry Manipulations of mutants & genotypes Genetic modifications Mutations naturally occurred or artificially-introduced 59 Genetic Modifications. m in N O R M A L IZE D PE A KA R E A NORM A LIZE D PEAK AR E A 0. Our Approaches To identify and screen novel starches from seeds Single kernel sampling (SKS) for starch analysis ‰ A methodology is established to screen novel starches from individual maize kernels while maintaining the vitality of each kernel ‰ SKS starch analysis may be conducted with high throughput when FACE (fluorophore-assisted carbohydrate electrophoresis) is used ‰ SKS may allow us to screen starch mutants from a large pool of seeds collected from natural plants or generated using artificiallyinduced mutagenesis 60 Genetic Modifications.I. V A L U E HPSEC RETENTIO N TIME. DP NORM A LIZE D DR IV AL UE 3000 2500 2000 1500 1000 500 0 10 12 14 16 18 20 b y MKI by S K S 1000 100 50 20 10 5 N OR M A L IZ E DD .58 Genetic Modifications. 2002.R .04 0. Our Approaches Single kernel sampling (SKS) for starch analysis ‰ Analyzing starch structure while maintaining kernel vitality ‰ Potential high throughput endosperm starch screening DEGREE OF P OLYMERIZATION. Cereal Chemistry.

characterize the activity profiles of starch biosynthetic enzymes and starch structure (e. and chain length Polydispersity Number average molecular weight Weight average molecular weight ‰ Starch chain length distribution ‰ Chain length distribution of starch ‰ Methods to characterize chain length distribution ‰ ‰ ‰ ‰ Size exclusion chromatography (SEC and HPSEC) Fluorophore-assisted carbohydrate electrophoresis (FACE) High performance anion exchange chromatography (HPAEC) Comparisons ‰ Structural parameters from chain length distribution ‰ Our capability to conduct carbohydrate analysis . without sacrificing starch yield 62 Starch Analysis Starch Fine Structure Analysis 63 Outline ‰ Why study starch fine structure? ‰ Fine structure and chain length distribution ‰ Introduction of molecular weight ‰ ‰ ‰ ‰ Molecular weight. degree of polymerization. and construct a database ‰ For a starch structure required by a desirable function.g. elucidate their relationships.61 Genetic Modifications. Our Approaches Roadmap to effective genetic starch modifications ‰ For selected genotypes. AND. amylopectin branching pattern) ‰ Correlate enzyme activity profile with starch structure. propose feasible enzyme activity profiles based on the database and certain hypotheses ‰ Create genotypes rendering proposed enzyme activity profiles.

stability. gel strength.g. viscosity.64 Fine Structure and Chain Length Distribution Why study “starch fine structure”? ‰ To differentiate starches (and starch derivatives) at molecular levels ‰ To define and document unique starches or starch derivatives (e. water adsorption. maltodextrins) in both research reports and patents ‰ To monitor product profiles and maintain product consistency ‰ To control important properties (digestibility. Carbohydrate Polymers. 2000. retrogradation. hazing. solubility. 43: 223-239 Structural parameters constructed 66 Fine Structure and Chain Length Distribution Chain length distribution is based on several basic concepts of molecular weight of polymers ‰ Molecular weight and degree of polymerization ‰ Polydispersity ‰ Number average molecular weight ‰ Weight average molecular weight . etc) of starches and starch derivatives ‰ To define goals for starch modifications ‰ To guide new product development 65 Fine Structure and Chain Length Distribution Starch fine structure is usually characterized by chain length distribution using cluster model Starch materials Specific treatments Completely debranched Chain length distribution characterized Chains are non-randomly clustered Thompson.

is used to quantify molecular weight of starch chains 68 Molecular Weight Polydisperse and polydispersity GGGGGGGGG GGGGG GGGGGGGGGGGGG This collection of molecules is polydisperse Polydispersity index (PDI): ‰ Ratio of the weight average molecular weight to the number average molecular weight ‰ Indicating the overall distribution of individual molecular weight in a batch of polymers ‰ PDI equal to 1 indicates only one length of polymer is present (e. expressed in DP. degree of polymerization.67 Molecular Weight Molecular weight. relative to the “unified atomic mass unit” (equal to 1/12 the mass of one atom of carbon-12) ‰ Degree of polymerization (DP): ‰ The number of monomeric residues in a polymer molecule ‰ “Chain length”.g. and colligative properties ‰ MN is more weighted by the small molecules in the molecular population . starch). 69 Molecular Weight Number average molecular weight (MN) ‰ MN = ΣiNiMi/ΣiNi . where Ni is the number of molecules of molecular weight Mi ‰ Can be determined by osmometry. PDI may vary significantly. end group titration.g. crystallized protein) ‰ In polysaccharides (e. and chain length ‰ Molecular weight (MW): ‰ The mass of one molecule of the substance.

000 101.000/1100 = 91. and sedimentation velocity ‰ MW is more weighted by the large molecules 71 Molecular Weight Polydispersity index = MW/MN 72 Molecular Weight An example of MW.000 1.100 Ni(Mi)2 100.000 100 1. X-ray scattering.67 . and polydispersity index Mi (DP) 100 10 Σ MW = ΣiNi(Mi)2/ΣiNiMi MN = ΣiNiMi/ΣiNi Ni 10 10 20 NiMi 1. small angle neutron scattering (SANS).8 (DP) 1.MN.70 Molecular Weight Weight average molecular weight (MW) MW = ΣiNi(Mi)2/ΣiNiMi. where Ni is the number of molecules of molecular weight Mi ‰ ‰ Can be determined by light scattering.100/20 = 55 (DP) So.000 101.8/55 = 1. polydispersity index = 91.

but not always. to differentiate starches at molecular level ‰ Starch molecules need to be debranched to release chains 75 Chain Length Distribution A procedure to prepare debranched starch materials Starch extraction Dispersed in DMSO Adding buffer + isoamylase Incubation Moisture evaporated Debranched starch in DMSO Waxy starches do not need AP isolation Amylopectin isolated Adding buffer + isoamylase Incubation Moisture evaporated Debranched AP in DMSO Adding buffer + beta-amylase Incubation Beta-limit dextrin (BLD) extraction Re-dispersion in DMSO Adding buffer + isoamylase Incubation Adding pullulanase Incubation Pullulanase needed to remove short stubs (DP2) Debranched BLD in DMSO Moisture evaporated .73 Chain Length Distribution Now. MN. the chain length profiles are compared among different starches. the data of chain length distribution are used to calculate structural parameters of starch or AP clusters 74 Chain Length Distribution Chain length distribution of normal and ae corn starch ‰ These figures (by HPSEC) are called “chain length distribution (profile)” ‰ MW. e. alpha-limit dextrin ‰ Then. let us discuss chain length distribution ‰ Chain length distribution of following materials have been characterized ‰ Starch (containing both amylose and amylopectin) ‰ Isolated amylopectin ‰ Beta-limit dextrin of amylopectin ‰ Other starch derivatives. and polydispersity index can be calculated from the curve ‰ Usually.g.

76 Chain Length Distribution Analysis of chain length distribution ‰ Three types of debranched starch material are usually prepared ‰ Debranched starch ‰ Debranched amylopectin ‰ Debranched beta-limit dextrin ‰ Three types of separation methods are often used to describe the chain length distribution of these materials ‰ Size exclusion chromatography (SEC or HPSEC) ‰ Fluorophore-assisted carbohydrate electrophoresis (FACE) ‰ Anion exchange chromatography (HPAEC) 77 Chain Length Distribution Size exclusion chromatography (SEC or HPSEC) By Dr.forumsci.co. Shulamit Levin. http://www.il/HPLC/modes/modes14.htm 78 Chain Length Distribution ‰ Eluent from HPSEC columns passes through a refractive index (RI) detector for quantifying the mass of carbohydrate molecules ‰ Molecular standards are used to calibrate columns and determine the molecular weight of samples ‰ Eluent may pass through a multiple angle laser light scattering (MALLS) detector for molecular weight determinations .

forumsci. 80 Chain Length Distribution Fluorophore-assisted carbohydrate electrophoresis (FACE): labeling of oligosaccharide using 1-aminopyrene3.il/HPLC/modes/modes15. Shulamit Levin. http://www.79 Chain Length Distribution Calibration curve and MW distribution of SEC By Dr.htm.6.co.8-trisulfonate (APTS) - O 3S NH2 NaBH 3 CN NH SO 3 - + - O 3S SO 3 - - O 3S SO 3 - Oligosaccharide APTS APTS adducts Excitation 488 nm Emission 520 nm Courtesy of Beckman-Coulter 81 Chain Length Distribution APTS adducts of carbohydrate molecules has 2 properties NH SO3 - - O3S SO3 - ‰ The APTS adducts are negatively charged. so may migrate in an electric field of electrophoresis ‰ The APTS adducts may release detectable fluorescent emission with laser excitation at 488 nm .

82 Chain Length Distribution Molecular weight is determined by migration time Number of molecules is determined by fluorescent signal NH SO3 - Longer migration time due to larger CHO molecule Weaker fluorescent signal due to fewer molecules - O3S NH SO3 SO3 - - - O3S SO3 - NH SO3 - NH SO3 - Shorter migration time due to smaller CHO molecule Stronger fluorescent signal due to more molecules - O3S NH SO3 - - O3S NH SO3 - SO3 - SO3 - - O3S SO3 - O3S SO3 - 83 Chain Length Distribution FACE conducted using capillary electrophoresis with laser-induced fluorescence 3 1 4 RFU 4 Detector 2 3 TIME La se r Electrophoresis 2 1 Courtesy of Beckman-Coulter 84 Chain Length Distribution Electrophoregram of FACE FU 3 APTS 30 2 10 40 50 80 1 20 0 0 10 20 30 Courtesy of Beckman-Coulter .

the higher the salt concentration required for desorption Courtesy of Amersham Biosciences . negatively charged Negatively charged sample molecules adsorbed by stationary phase may be desorbed by mobile phase Courtesy of Amersham Biosciences 87 Chain Length Distribution The adsorption/desorption equilibrium is governed by: ‰ The amount of negative charge of sample molecules ‰ The ionic strength of mobile phase The greater the net charge. the stronger the adsorption The greater the net charge. different types of charge or neutral Stationary phase.85 Chain Length Distribution Comparison between HPSEC and FACE HPSEC FACE 86 Chain Length Distribution High performance anion-exchange chromatography equipped with pulsed amperometric detector (HPAEC-PAD) Sample molecules. positively charged Mobile phase.

which may be affected by pH ‰ Elution power.88 Chain Length Distribution The retention time of a molecule is determined by ‰ Its net negative charge. carbohydrates are negatively charged ‰ Thus. there exists an adsorption/desorption equilibrium of carbohydrate molecules with stationary phase ‰ The higher DP of carbohydrate molecule. carbohydrate molecules with different DP are eluted at different retention time and separated 90 Chain Length Distribution PAD catalyzes the electrooxidation of carbohydrate molecules in high pH solutions. which is controlled by gradient elution Courtesy of Amersham Biosciences 89 Chain Length Distribution Carbohydrates can be separated via anion-exchange ‰ In basic solution (high pH). and gives signals proportional to the amount of molecules Oxidation Reduction Typical waveform for PAD in alkaline solution at a gold working electrode (Courtesy of Dionex Corporation) . the greater net negative charge it possesses. the higher ionic strength (salt concentration) needed to desorb the molecule ‰ Using gradient eluent.

100 0. Carbohydrate Research.91 Chain Length Distribution HPAEC-PAD gives a chromatogram similar to FACE 0.003 0 25 50 75 100 125 150 175 min 200 Chain length distribution of debranched amylopectin of sorghum starch by HPAEC-PAD 92 Chain Length Distribution Pros and cons of HPSEC. 2005.125 0. but unable to (with current techniques) give a chain length profile with broader DP range ‰ HPAEC is similar to FACE 93 Chain Length Distribution HPSEC and FACE (possible HPAEC) are complementary We have successfully demonstrated the compatibility of HPSEC and FACE in starch analysis. FACE.160 0.050 0. 340: 701-710 .000) but with relatively low resolution >DP5 ‰ FACE provides baseline resolution up to DP100.025 -0.075 0. and established an HPSEC-FACE methodology balancing both high resolution and broad DP range Yao et al. and HPAEC ‰ HPSEC provides information of a broad range of molecular weight (>DP10.

chains connecting three clusters B1a chain.I 60 40 20 0 10 12 14 16 18 20 B3 ICL + 2. 136: 3515–3523 .5 DP) ae wx Cluster repeat distance (26.2 DP) B2 chain.5 DP) wx wx β-limit dextrin B chain stub 1 or 2 DP A chain stub 2 or 3 DP ICL of B1-a (3. long intra-cluster chains A chains Yao et al. DP ae wx wx RELATIVE MOLAR AMOUNT DEGREE of POLYMERIZATION. then what? How to calculate structural parameters of starch or amylopectin? ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ Number average chain length Average external chain length Distribution of external chain length Average internal chain length Distribution of internal chain length Average number of branches per cluster Average cluster repeating distance Distance of adjacent branches Chain length distribution of B2 chains Chain length distribution of B3 chains Chain length distribution of long B1 chains Chain length distribution of short B1 chains 95 Chain Length Distribution A comparison between wx and ae wx starch Debranched amylopectin DEGREE of POLYMERIZATION. short intra-cluster chains B1b chain. chains connecting two clusters B3 chain. DP 1000 100 50 20 10 3 Debranched β-limit dextrin DEGREE of POLYMERIZATION.94 Chain Length Distribution OK.5 0 10 12 14 16 18 20 DEGREE of POLYMERIZATION. we got chain length distribution.5 40 20 B1-a ICL+ 2.5 B1-b ICL+ 2. DP 60 NORMALIZED D. 2004. Plant Physiology.I 1000 100 50 B3 20 10 3 NORMALIZED D. min 96 Chain Length Distribution Decoding chain length distributions ICL of B3 (53. DP RELATIVE MOLAR AMOUNT 1000 100 50 20 10 3 5 4 3 2 1 0 10 12 14 16 18 20 RETENTION TIME.5 B2 B1-b B1-a A B2 ICL+ 2.R. min 15 1000 100 50 B3 B2 20 10 B1-b B1-a 3 A Average CL 10 A chain amount Cluster amount 5 B1-a ICL+ 2.R.5 0 10 12 14 16 18 20 RETENTION TIME.5 DP) Average number of branches per cluster (ANBPC) ICL of B2 (26.

the granules may absorb limited amount of water ‰ Generally.97 Capability of Carbohydrate Analysis ‰ What do we have? ‰ ‰ ‰ ‰ Beckman P/ACE MDQ capillary electrophoresis with LIF to conduct FACE Waters HPLC system to conduct HPSEC and many other separations Dionex HPAEC for sugar and starch analysis Other systems ‰ What can we do? ‰ Analysis of starch structure ‰ Analysis of starch derivatives and hydrolysates ‰ Analysis of simple sugars. insoluble in water ‰ When put in cold water. and polysaccharides ‰ Throughput? ‰ FACE up to 50~80 samples/day ‰ HPSEC up to 10~20 samples/day ‰ HPAEC up to 10 samples/day ‰ Beyond the throughput? ‰ ‰ ‰ ‰ Methodology development Product characterization Optimize & define industrial process And much more…… 98 Functional Properties of Starch ‰ Thermal properties ‰ Gelatinization ‰ Swelling power ‰ Retrogradation ‰ Glass transition temperature ‰ Rheological properties ‰ Brabender ‰ RVA (rapid visco analyzer) ‰ Viscoelasticity 99 Thermal Properties Starch gelatinization ‰ Isolated starch is in granular form. oligosaccharides. the swelling is reversible . when temperature is below a certain value (for most around 60°C).

“Gelatinization” begins ‰ The exact temperature dependent on the specific starch. it is a good indicator of the initial gelatinization temperature of a given starch .g.100 Thermal Properties Starch gelatinization ‰ With higher temperatures (usually >65°C) irreversible swelling of starch granules may occur. which can be quantified by DSC 102 Thermal Properties Starch gelatinization ‰ Since the loss of birefringence occurs at the time of initial gelatinization. granule sizes. e. plant species. etc 101 Thermal Properties Starch gelatinization Peak temperature Endothermic heat flow Onset temperature Temperature ‰ Gelatinization occurs in a range of temperature. mutants. cultivars.

divided by the weight of the original dry starch 105 Thermal Properties Starch retrogradation ‰ Irreversible insolubilization of starch paste with formation of a precipitate or gel depending on concentration ‰ Also known as "set back" ‰ Retrogradation may be caused by amylose as in regular starches ‰ Retrogradation may also be caused by amylopectin as in waxy starches ‰ Adjacent linear chains of amylose or amylopectin form double helices via hydrogen bonds and further aggregates to form crystalline regions . the granules undergoes irreversible swelling The degree is a function of the starch type and the presence of any physical or chemical modification The swelling power is a measure of the hydration capacity of the starch and is expressed as the weight of centrifuged swollen granules.103 Thermal Properties Starch gelatinization 65C 70C 85C ‰ The gelatinization range refers to the temperature range over which all the granules are fully swollen ‰ Increased temperature and shear may cause granule disruptions ‰ Increased translucency and increased viscosity may be observed during gelatinization 104 Thermal Properties Swelling power of starch granules Swelling power = weight-1/weight-2 Swelling Centrifugation Weight-1 Weight-2 ‰ ‰ ‰ When an aqueous suspension of starch is gelatinized.

1998. primarily caused by amylopectin retrogradation ‰ Ageing of cooked rice and tortillas ‰ Syneresis of starch paste after freeze-thaw recycles ‰ Opaqueness development of starch paste/gel/dispersion ‰ And other “bad” things So it’s a reason for starch substitution to make stabilized starch 107 Thermal Properties Starch retrogradation is also associated with… ‰ Production of resistant starch ($1. Cereal Chemistry 75 (6) .5/lb) ‰ Preparation of slowly digesting starch ‰ Film formation by amylose ‰ And possibly other “good” things So it’s a reason to intensify starch retrogradation for value-added products 108 Thermal Properties Starch retrogradation studied using DSC Liu and Thompson.106 Thermal Properties Starch retrogradation is associated with… ‰ Bread staling.

Cereal Chemistry 75 (6) 110 Thermal Properties Starch retrogradation studied using DSC Liu and Thompson. Cereal Chemistry 75 (6) 111 Thermal Properties Starch retrogradation studied using DSC Liu and Thompson.109 Thermal Properties Starch retrogradation studied using DSC Liu and Thompson. 1998. Cereal Chemistry 75 (6) . 1998. 1998.

etc 114 Rheological Properties Brabender Viscograph Cooled paste stability Peak viscosity Setback Breakdown .112 Thermal Properties How to manipulate starch retrogradation??? ‰ Chemical modifications. etc to affect retrogradation. substitution (stabilization) ‰ Enzymatic modifications. pastes. e. lipids. suspensions. using partial beta-amylolysis ‰ Genetic modifications. e.g. su2 wx double mutants You may also add sugars. but structure modifications are the most effective approaches 113 Rheological Properties Brabender Viscograph ‰ Gelatinization of starch and materials containing starch ‰ Hot and cold viscosity ‰ Stability of thickening agents or binders ‰ Acid stability of starch ‰ Measurement of liquids.g.

oils Pie filling. gum confectionery Confectionery. soups. salad dressings Emulsions. suspension. formulated liquid foods Formulated foods. stabilizing.115 Modified Food Starches ‰ Converted starches ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ ‰ Acid conversions Oxidized starches Pyroconversions or dextrinizations Distarch phosphate Distarch adipate Starch acetate Starch phosphate Starch sodium octenyl succinate Hydroxypropylated starch Beta-amylase treatment to reduce retrogradation Pregelatinization for cold water-swelling starch ‰ Cross-linked starches ‰ Stabilized starches ‰ Other modifications 116 Modified Food Starches Properties and applications of modified starches Process Acid conversion Oxidation Dextrins Cross-linking Esterification/ Etherification Pregelatinization Dual modifications Function/property Viscosity lowering Adhesion. encapsulation. Yellow. flavorings. stable viscosity on storage of the paste ‰ Dextrin ‰ Starch hydrolysis products obtained in a dry roasting process either using starch alone or with trace levels of acid catalyst ‰ Dextrins are characterized by good solubility in water to give stable viscosities. breads. baking (gloss). ‰ Oxidized starch ‰ Starch suspension is usually treated with sodium hypochlorite ‰ Commercial oxidized starch is granular and is insoluble in cold water ‰ It is characterized by a whiter color than native starch. British Gums and Solution-stable Dextrins . texturizing Stabilization. gelling Binding. filtration. Acid-thinned starches tend to have a much lower hot viscosity than native starch and a strong tendency to gel when cooled. increased paste clarity and a low. and drying once the desired degree of conversion is reached ‰ This results in a reduction in the average molecular size of the starch polymers. infant foods. spices. batters. low temperature storage Cold water swelling Combinations of properties Application Gum candies. frozen foods Premix As you can imagine… 117 Modified Food Starches Converted starches ‰ To reduce the viscosity and swelling power and increase the concentration in the dispersions ‰ Acid-thinned starch: ‰ Starch suspension is treated with dilute acid at a temperature below the gelatinization point ‰ Granular form of the starch is maintained and the reaction is ended by neutralization. soups. Four types exist: White. high solubility Thickening. coating. bakeries. puddings.

g. and shear forces ‰ Cross-linking is widely used to prepare chemically-modified starches for the processed food industry ‰ Distarch phosphate ‰ A starch cross-linked with a phosphate linkage. from adipic acid ‰ Most cross-linked food starches contain less than one crosslink per 1000 glucopyranosyl units 119 Modified Food Starches Starch stabilization ‰ Starch stabilization ‰ “stabilization” is sometimes used to indicate the presence of a monofunctional chemical substituent which has the effect of stabilizing paste viscosity ‰ Stabilized modified starches may be hydroxypropyl or carboxymethyl starch ethers ‰ The monofunctional substituents also can be phosphate or acetyl ester groups ‰ Generally the D. The substituent groups have the effect of providing steric hindrance to chain association which stabilizes viscosity by preventing possible retrogradation. heat treatment. 120 Modified Food Starches Starch alkenyl succinate ‰ Starch alkenyl succinate ‰ A chemically modified starch produced by treating starch with alkenyl succinic anhydride under controlled pH conditions ‰ Commercial alkenyl succinic anhydride available for use in food is the octenyl form ‰ These starches have lipophilic ("oil-loving") properties and are used in emulsions and encapsulation ‰ Starch octenyl succinate ‰ Common name given to Starch n-Octenyl succinate ‰ Made by treating starch with n-Octenyl succinic anhydride at pH 8-8.S.01 and 0. (degree of substitution) of these starches is between 0.2.118 Modified Food Starches Cross-linked starch ‰ Cross-linked (bonded) starch ‰ Starch is treated with bi-functional reagents so that a small number of the starch polymer chains are chemically linked by the cross linking reagent ‰ Cross-linking inhibits granule swelling on gelatinization and gives increased stability to acid.5 ‰ Anionic due to a carboxyl group and hydrophobic due to the C8 unsaturated alkene chain ‰ Food uses include encapsulation of flavors and emulsion stabilization . e. from reagents such as sodium trimetaphosphate or phosphorus oxychloride ‰ Distarch adipate ‰ A starch cross-linked with a adipate linkage.

and their hydrolysates ‰ Isoamylase and pullulanase are debranching enzymes with different functional behaviors ‰ Widely used in starch structure analysis ‰ Widely used in producing dextrose and maltose .4)-linkages of glucan chains of amylose and amylopectin ‰ Converts starch + H2O to maltodextrins or syrups comprising simple sugars and oligo or polysaccharides ‰ Average extent of hydrolysis can be quantified using DE value (dextrose equivalent) ‰ Products with DE<20 is defined as maltodextrin.4) and α-D-(1. pregelled starch. DE>20 is defined as (corn) syrup ‰ β-Amylase ‰ Exoenzyme cleaves two glucosidic units sequentially from non-reducing ends of glucan chains ‰ Exhaustive β-amylolysis converts starch to maltose + β-limit dextrins ‰ Partial β-amylolysis was used to shorten amylopectin external chains to retard retrogradation 123 Starch Enzymes and Starch Digestibility Starch hydrolases ‰ Amyloglucosidase (glucoamylase) ‰ Exoenzyme that digests ungelatinzied. or fully gelatinized starch ‰ Releasing glucose by hydrolyzing both α-D-(1. dispersion and texture 122 Starch Enzymes and Starch Digestibility Starch hydrolases ‰ α-Amylase ‰ Endoenzyme cleaves internal α-D-(1.121 Modified Food Starches Pregelatinized starch ‰ A type of starch which has been gelatinized and dried by the manufacturer before sale to the customer in a powdered form ‰ Pregelatinized starch can be made by drum drying. or extrusion from either native or modified starch ‰ Pregelatinized starch develop viscosity when dispersed in cold or warm water without the need for further heating ‰ Pre-gelatinized starch is also known as precooked starch.g. amylopectin. cold water soluble starch.6)-linkages of branched α-D-glucan. or cold water swelling starch (CWS) ‰ The degree of granular integrity and particle size have a major influence on their properties. spray drying. instant starch.6)-linkages from non-reducing ends ‰ Industrially converts starch or liquefied starch (after α-amylase hydrolysis) to dextrose (glucose) ‰ Debranching enzymes ‰ Specifically hydrolyze the α-D-(1. partially gelatinized. e. e.g. glycogen.

4) and α-D-(1.6)-linkages to produce glucose for absorption ‰ Other carbohydrate–digesting enzymes ‰ Sucrase & Lactase Adapted from Dr. resulting from the physical structure of retrograded starch molecules after the starch granules are gelatinized ‰ Type IV.124 Starch Enzymes and Starch Digestibility Starch digestion ‰ Starch is digested in the human body ‰ Firstly by α-amylase (salivary and pancreatic) ‰ Yields maltose. maltotriose. It hydrolyzes α-D-(1. it may be fermented in the large intestine by colonic microflora ‰ RS has been classified in four different categories ‰ Type I. resulting from the physical structure of the uncooked. and α-dextrins ‰ Maltose and maltotriose are hydrolyzed to glucose by maltase ‰ α-dextrin by α-dextrinase (intestinal brush border enzymes) ‰ α-Dextrinase (glucoamylase) is found at the brush border of the small intestines. cross-linking) that interferes with the enzyme digestion. or “resistant starch” is usually readily digested in the proximal colon by bacterial amylases. Hamaker’s lecture in Starch Short Course 126 Starch Enzymes and Starch Digestibility Resistant starch ‰ Resistant starch (RS) ‰ Starch that is resistant to digestion by α-amylase ‰ Defined as "the sum of starch and products of starch degradation not absorbed in the small intestine of healthy individuals. lactose make up “net carbs” ‰ Fiber constituents are not digested by the body’s enzymes ‰ Starch undigested by host enzymes. and is about 20% utilized as energy for the body Adapted from Dr. native starch granules ‰ Type III." ‰ While RS escapes digestion in the small intestine. . maltodextrin. Hamaker’s lecture in Starch Short Course 125 Starch Enzymes and Starch Digestibility Starch digestion ‰ Starch. resulting from physical inaccessibility in intact tissues or other large particulate materials ‰ Type II. sucrose. glucose. resulting from chemical modification (e.g.

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