| HS Code | 679601 |
| Product Name | Alpha-Amylase |
| Cas Number | 9000-90-2 |
| Ec Number | 3.2.1.1 |
| Source | Bacillus subtilis / Aspergillus oryzae |
| Appearance | Lyophilized powder or clear to amber liquid |
| Molecular Weight | ~55 kDa (varies by source) |
| Optimal Temperature | 50–60 °C |
| Optimal Ph | 6.0–7.0 |
| Specific Activity | ≥500 U/mg (varies by preparation) |
| Solubility | Soluble in water; insoluble in ethanol |
| Storage Conditions | Store at 2–8 °C, airtight, protected from moisture |
| Inhibitors | EDTA, urea, heavy metal ions |
| Activators | Calcium ions (Ca²⁺) |
As an accredited Alpha-Amylase factory, we enforce strict quality protocols—every batch undergoes rigorous testing to ensure consistent efficacy and safety standards.
| Packing | Alpha-Amylase, 1 kg, supplied in a sealed polyethylene bag inside a fiber drum, with desiccant, labeled for lab use. |
| Container Loading (20′ FCL) | 20′ FCL: load Alpha-Amylase drums securely, restrain cargo, label clearly, avoid contamination, ensure ventilation, and protect from heat/moisture. |
| Shipping | Alpha-Amylase is shipped as a temperature-sensitive enzyme, typically refrigerated or frozen to preserve activity. Packaging uses moisture-resistant, airtight containers to prevent degradation. Non-hazardous under normal conditions, but avoid heat and humidity. Transport via cold-chain couriers ensures stability, with documentation noting handling requirements and shelf-life restrictions upon delivery. |
| Storage | Store Alpha-Amylase at 2–8°C in a tightly sealed, desiccated container, protected from moisture and light. Lyophilized powder is generally stable under these conditions. For solutions, follow manufacturer recommendations, often at 2–8°C, and avoid repeated freeze-thaw cycles to preserve enzymatic activity. |
| Shelf Life | Alpha-Amylase typically has a shelf life of up to one year when stored at 2–8°C, protected from moisture. |
In corn wet milling, potato starch, and tapioca starch refineries, a thermostable bacterial alpha-amylase (EC 3.2.1.1) is metered into starch slurry immediately upstream of the primary jet cooker. The slurry is maintained at 30–35% dry solids, pH 5.8–6.2, and free calcium 40–80 mg/kg; calcium concentration is the dominant kinetic variable because the enzyme active site requires a conserved Ca²⁺-binding structure. Slurry is pumped through a steam jet cooker at 103–107°C for 5–8 min, then flashed to 95–97°C and held in a continuous stirred liquefaction column for 90–120 min. Dose rates for a 240 KNU/g liquid preparation range from 0.35 L to 0.80 L per metric tonne of dry substance, depending on dextrose equivalent target and the alpha-1,6 branch content of the starch source. A DE of 10–14 after liquefaction is typical before pH is reduced to 4.2–4.5 for saccharification with glucoamylase at 60°C for 48–72 h, producing a glucose-rich hydrolysate of DX 95–96. Terminal products from this route include glucose syrup, high fructose corn syrup, maltodextrin, and crystalline dextrose. Food-grade enzyme compliance is referenced to the JECFA alpha-amylase monograph, FCC 15th ed. enzyme specifications, and Regulation (EC) No 1332/2008. Thermal over-holding above 107°C or free calcium below 30 mg/kg produces colour and 5-hydroxymethylfurfural formation through Maillard reactions; plant operations that recycle evaporator condensate can shift ionic strength and require dosage revalidation against the target DE curve.
| Parameter | Wet-mill sweetener liquefaction | Dry-grind ethanol liquefaction |
|---|---|---|
| Dry solids | 30–35% | 28–34% |
| pH window | 5.8–6.2 | 5.5–6.0 |
| Free calcium | 40–80 mg/kg | 50–120 mg/kg adjusted |
| Jet cooking temperature | 103–107°C | 103–105°C |
| Holding temperature/time | 95–97°C, 90–120 min | 95–97°C, 90–180 min |
| Dose rate, 240 KNU/g | 0.35–0.80 L/t dry solids | 0.40–0.80 L/t corn |
| Target DE after liquefaction | 10–14 | 6–10 |
| Primary finished-product standard | FCC 15th ed., JECFA, Regulation (EC) No 1332/2008 | ASTM D4806-21 for denatured fuel ethanol |
Where whole maize or sorghum is milled through a hammer mill to a geometric mean particle size of 2–3 mm, the resulting meal is slurried with recycled thin stillage and fresh water to 28–34% dry solids before a thermostable alpha-amylase is added at 0.40–0.80 L/t corn for a 240 KNU/g preparation. The slurry is heated in a pre-mix tank to 83–88°C, passed through a jet cooker at 103–105°C, and held at 95–97°C for 90–180 min; the target DE of 6–10 is below the sweetener liquefaction range because excessive dextrose formation before fermentation can increase osmotic stress and reduce yeast viability. After liquefaction, pH is reduced to 4.5–5.0, glucoamylase and yeast are added for simultaneous saccharification and fermentation at 32–34°C, and the resulting beer is distilled to produce fuel ethanol and distillers’ grains with solubles. Finished ethanol must meet ASTM D4806-21 for denatured fuel ethanol, while the enzyme preparation is supplied under the JECFA alpha-amylase monograph and Regulation (EC) No 1332/2008 where food-grade host strains are used. The main process conflict in dry-grind liquefaction is retrogradation of starch-lipid complexes in recycled stillage; pH depression below 5.2 from lactic acid in backset or calcium chelation by phytate hydrolysis reduces thermostability and requires either calcium mineral adjustment or enzyme dose elevation. Published data for full-plant ionic-strength interactions remain limited, but liquefaction failures on production lines are commonly observed as viscosity rebound after the hold tank and as elevated glucan retention in DWG.
The separate cereal cooker receives rice, maize, or sorghum grits at 25–45% of total grist, with a high-temperature bacterial alpha-amylase added at 0.10–0.25 kg/t adjunct dry weight. The adjunct is slurried at 65–70°C, ramped to 95–100°C, and held for 30–60 min until the starch is liquefied and iodine staining is reduced. The cooked adjunct is then combined with the main malt mash at 62–65°C, where the thermostable alpha-amylase activity is largely inactivated by malt beta-amylase conditions and the remaining hydrolysis proceeds through malt diastase. Terminal products include high-adjunct lager, dry stout, and sorghum-based gluten-free beer. Brewery compliance draws on the ASBC and Analytica-EBC wort extract methods, with the enzyme preparation itself falling under JECFA, FCC 15th ed., and Regulation (EC) No 1332/2008 as a processing aid. The main process limit is foam stability: overdosing alpha-amylase above 0.25 kg/t adjunct can generate low-molecular dextrins that do not support foam-positive protein foam matrices and can increase fermenter foaming due to accelerated carbohydrate release.
Formulated as a fungal alpha-amylase preparation of 2500 FAU/g from Aspergillus oryzae, the enzyme is metered into wheat flour at 10–50 ppm flour weight during dough mixing at 20–28°C. It converts damaged starch granules into maltose and maltotriose during proofing and is inactivated above 70°C in the crumb. Published data for this specific configuration are limited to comparative crumb firmness rather than absolute enzyme-unit input, but production trials indicate that thermostable bacterial alpha-amylase overdosing above 50 ppm can produce a dextrin-rich, gummy crumb. Terminal products are pan bread, buns, and frozen dough. Food enzyme compliance is governed by the JECFA alpha-amylase monograph, FCC 15th ed., and Regulation (EC) No 1332/2008; most jurisdictions classify fungal alpha-amylase as a processing aid requiring no final ingredient declaration.
Continuous enzymatic desizing of starch-sized cotton and cotton-polyester warps requires a pad-batch or pad-steam range with controlled wet pick-up and wash modules. A bacterial alpha-amylase preparation at 30,000 MWU/g is dosed at 1.0–4.0 g/L in the saturator, together with 0.5–1.0 g/L nonionic wetting agent, at pH 6.0–7.0 and 60–80°C. The fabric is batch-rolled for 8–16 h or steamed for 30–60 min, followed by a wash module at 85–95°C to remove dextrin hydrolysates. Desizing efficiency is graded by the TEGEWA violet scale, with readings of 6–9 typically required before reactive dyeing; residual starch above this level produces uneven dye uptake and can cause batch-to-batch shade variance. Terminal products include desized greige fabric for direct printing, reactive dyeing, and resin finishing. Regulatory compliance is established through REACH registration of the enzyme preparation, user obligations under the ZDHC Manufacturing Restricted Substances List where applicable, and the supplier safety data sheet. Bath limitations include iron above 0.2 mg/L and pH above 8.0, which reduce activity and may require chelating agents or process-water softening before the pad liquor is prepared.
Size press starch viscosity is controlled by adding a thermostable alpha-amylase to native corn starch slurry after the starch has been jet-cooked at 95–105°C and cooled to 75–80°C; the enzyme is typically applied at 0.1–0.5% of starch dry weight. The reaction is held for 10–30 min until a Brookfield viscosity of 30–80 mPa·s is reached at 60°C, then the enzyme is deactivated by heating above 95°C for 5–10 min to prevent viscosity drift in the size press run tank. The resulting carrier starch is fed to a size press on fine paper or to a corrugator as part of a Stein-Hall corrugating adhesive. Terminal products include surface-sized coated paper, laminated board, and corrugated containers. Compliance for indirect food contact is established through FDA 21 CFR 176.170; paperboard performance is measured by ISO 535 Cobb water absorption and TAPPI T 419 starch content where specified. Overdosage beyond 0.5% starch dry weight reduces starch molecular weight to a level that lowers surface strength and corrugator water resistance, and batch-to-batch variation in native starch paste viscosity makes trial runs on the target size press necessary before setting the final enzyme dose.
Automatic dishwashing detergent tablets and powders carry alpha-amylase as a coated granulate at 0.5–2.0% by weight of the finished product, where the enzyme must survive alkaline pH 9.0–10.5, bleach systems, and calcium-sequestering builders. The granulate is dry-blended after spray-drying to avoid activity loss above 55°C; in the main wash cycle at 40–60°C, the enzyme hydrolyzes starch films on ceramic and stainless steel surfaces before the rinse phase. Terminal products include phosphate-free automatic dishwashing tablets, gel packs, and industrial warewash detergents. Detergent enzyme stability is evaluated under the A.I.S.E. Charter and Regulation (EC) No 648/2004; cleaning performance is assessed by industry-relevant mechanical dishwashing protocols rather than a single ISO endpoint. The main incompatibility is the combination of strong calcium chelators such as methylglycine diacetic acid and citrate at high builder concentrations, which removes the free calcium required for alpha-amylase conformational stability in the wash bath; granulate coatings must therefore be formulated with calcium salts to preserve thermostability during storage and release.
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Alpha-Amylase (EC 3.2.1.1) is an endo-acting calcium metalloenzyme that randomly hydrolyzes internal α-1,4-D-glucosidic linkages in amylose and amylopectin. The product described here is a thermostable liquid preparation derived from Bacillus licheniformis, supplied under the model designation Alpha-Amylase 120L as a brown to amber aqueous concentrate standardized to 120 KNU(T)/g with an activity tolerance of ±5%. The liquid has density 1.20–1.25 g/mL at 25 °C, pH 5.5–6.5, and is preserved for microbial stability. A lower-activity granulate grade, Alpha-Amylase 60G, is available at 60 KNU(T)/g for solid-phase detergent and textile operations. The preparation is produced under ISO 9001:2015 and conforms to the FCC enzyme preparation monograph and JECFA food enzyme specifications. The enzyme is not active on α-1,6 branch points, does not release glucose, and requires available Ca²⁺ for high-temperature operation. Storage is recommended at 0–25 °C in closed containers; freezing can cause protein aggregation and partial activity loss. Representative release criteria for the liquid food-grade product are provided in Table 1.
| Parameter | Test method | Release limit |
|---|---|---|
| Declared activity | Ceralpha α-amylase assay | 115–125 KNU(T)/g |
| Density at 25 °C | Oscillating U-tube density meter | 1.18–1.22 g/mL |
| pH | Potentiometric | 5.5–6.5 |
| Lead | ICP-MS, USP <233> | ≤5 mg/kg |
| Cadmium | ICP-MS, USP <233> | ≤0.5 mg/kg |
| Arsenic | ICP-MS, USP <233> | ≤3 mg/kg |
| Total aerobic count | ISO 4833-1 | ≤10,000 CFU/g |
| Escherichia coli | ISO 16649-2 | Absent in 25 g |
| Salmonella | ISO 6579 | Absent in 25 g |
| Storage stability | Supplier stability protocol | 0–25 °C, 18 months |
The random internal cleavage mode causes a sharp viscosity decrease before substantial reducing sugar formation. At the jet-cooker outlet, Brookfield viscosity at 60 °C typically drops from more than 1000 mPa·s to 50–150 mPa·s within 5–10 min, while dextrose equivalent remains below 5. By contrast, beta-amylase (EC 3.2.1.2) is an exo-acting enzyme that releases maltose from non-reducing chain ends but stops at α-1,6 branches. Glucoamylase (EC 3.2.1.3) is an exo-acting enzyme that releases β-D-glucose from both α-1,4 and α-1,6 linkages but is inactivated above 65 °C. Pullulanase (EC 3.2.1.41) cleaves α-1,6 branch points but does not efficiently attack long linear chains. Table 2 summarizes these specificity and operating differences.
| Enzyme | EC number | Mode | Linkage specificity | Principal products | pH optimum | Temperature optimum |
|---|---|---|---|---|---|---|
| Thermostable bacterial alpha-amylase | 3.2.1.1 | Endo | α-1,4 | Maltodextrins, oligosaccharides | 6.0–6.5 | 95–105 °C with Ca²⁺ |
| Beta-amylase | 3.2.1.2 | Exo | α-1,4 | Maltose | 4.5–5.5 | 55–65 °C |
| Glucoamylase | 3.2.1.3 | Exo | α-1,4 and α-1,6 | Glucose | 4.0–4.5 | 60–65 °C |
| Pullulanase | 3.2.1.41 | Debranching | α-1,6 | Linear oligosaccharides | 4.5–5.0 | 60–65 °C |
The practical consequence is that the thermostable alpha-amylase is specified upstream of saccharification because the exo-acting enzymes cannot survive jet-cooker temperatures. The product is also selected over fungal alpha-amylase when the process stream must remain at pH 6.0–6.5 and the target is a low-DE maltodextrin rather than a glucose or maltose syrup.
In continuous corn wet-milling, a starch slurry of 30–35 wt% dry solids is adjusted to pH 6.2 with sodium carbonate or dilute sodium hydroxide and dosed with 0.4–0.8 kg of the 120 KNU(T)/g liquid per metric ton of dry starch. The dosed slurry passes through a steam-injection jet cooker rated for 0.6–1.0 MPa steam pressure and a hold tube residence of 5–7 min at 105–110 °C. Calcium is maintained at 50–80 ppm Ca²⁺. At 105 °C, the random hydrolysis prevents gelation and reduces viscosity before the secondary hold. If free Ca²⁺ falls below 20 ppm, the observed production failure is not instantaneous pH collapse but a viscosity plateau in the secondary tank and partial starch retrogradation that increases differential pressure across downstream plate-and-frame filters.
Secondary liquefaction proceeds at 90–95 °C for 90–180 min. Progress is monitored by iodine color disappearance and by dextrose equivalent titration according to ISO 5377; the syrup is discharged at DE 8–15. HPLC analysis according to ISO 10504 shows maltotriose, maltotetraose, and higher oligosaccharides, with free glucose below 1 wt%. The low glucose content is a consequence of the endo mechanism: the product does not generate glucose directly and cannot hydrolyze α-1,6 branch points. The syrup is cooled to 60–65 °C, adjusted to pH 4.2–4.5, and supplemented with glucoamylase and pullulanase for glucose release. Acidification to below pH 4.0 before iodine-negative liquefaction causes irreversible denaturation of the alpha-amylase and leaves insoluble starch carryover into saccharification.
Activity is expressed in KNU(T) units rather than protein mass; the unit is defined by a supplier reference assay using soluble starch at fixed pH, temperature, and calcium concentration. It cannot be directly converted to maltogenic amylase units, bacterial amylase units, or fungal amylase units without a product-specific correlation. The liquid is standardized to the declared activity with glycerol or sorbitol, and the batch release tolerance of ±5% is the primary production control. The enzyme is inhibited by Cu²⁺ and Zn²⁺; exposure to hypochlorite oxidants or strong cationic flocculants should be avoided in dosing lines because enzyme-flocculant complexes can reduce activity and increase filter pressure drop.
Where woven cotton or viscose warps are sized with native or modified starch, the same endo-acting mechanism is applied in a continuous desizing range. The liquid enzyme is padded at 0.5–2.0 g/L with a nonionic wetting agent at 0.1–0.3 g/L, maintained at pH 6.5–7.0 and 80–90 °C, and the fabric passes through a steamer or J-box with dwell time of 3–15 min depending on fabric weight. The random cleavage reduces starch molecular weight and enables hot-water removal without strong alkaline hydrolysis. Desizing efficiency is checked with TEGEWA violet staining; production specifications commonly require less than 0.5 wt% residual starch before bleaching. The product is not effective on PVA, CMC, or acrylic size blends, so mixed-sized warps require additional oxidative or chemical desizing. In process water with less than 20 ppm CaCO₃-equivalent hardness, half-life in continuous equipment may decline unless calcium chloride is added.
In automatic dishwashing detergents, the enzyme is delivered as an encapsulated granulate rather than a liquid. Finished detergent matrices operate at pH 9–11 and contain sodium percarbonate or other bleaching agents; liquid alpha-amylase would be deactivated within minutes by the alkaline bleach system. The granulate is dosed at 0.1–0.5 wt% of the finished formula and is designed to release after the initial alkaline surge. Starch film removal is evaluated in a mechanical dishwasher under EN 50242; residual activity after accelerated storage at 37 °C and 70% relative humidity for 4 weeks is a common release criterion. The enzyme’s calcium requirement becomes a formulation constraint because builders such as sodium citrate and methylglycine diacetic acid sequester Ca²⁺; in fully softened water, starch film removal may fall below target unless release kinetics or calcium compensation are adjusted. Published data for this specific configuration is limited outside supplier application sheets.
In dry-grind fuel ethanol, the pH and temperature are moderated relative to wet-milling starch processing. The slurry is liquefied at pH 5.5–6.0 and 85–90 °C with 20–40 ppm Ca²⁺, then saccharified at pH 4.2–4.5 and 60–65 °C with glucoamylase. The lower pH reduces scaling in evaporators and distillation, but it narrows the alpha-amylase stability margin; if pH drops below 5.0 before complete liquefaction, iodine-positive starch persists and ethanol yield declines. The thermostable bacterial grade is therefore preferred over fungal alpha-amylase, which denatures during slurry heating.
In flour-based baking, the bacterial thermostable grade is not always the preferred alpha-amylase. Fungal alpha-amylase from Aspergillus oryzae is heat-labile and denatures at 60–75 °C during baking, limiting dextrinization to the early oven phase. The thermostable bacterial enzyme can survive baking and may produce overdextrinization and a gummy crumb if dosage is not tightly controlled. When the bacterial grade is used for crumb softening, application rates are 20–60 ppm on flour weight and require validation of residual activity after baking. Crumb firmness is measured according to AACC 74-09. The product also differs from maltogenic alpha-amylase (EC 3.2.1.133), which acts preferentially on amylopectin and is used for shelf-life extension without excessive dough weakening. Replacement of one alpha-amylase class with another without reformulation can alter crumb structure, loaf volume, and starch retrogradation kinetics.
