| HS Code | 320112 |
| Product Name | Beta-Amylase |
| Ec Number | 3.2.1.2 |
| Source | Barley malt or Bacillus cereus |
| Appearance | White to light brown powder |
| Optimum Temperature | 50-60 °C |
| Optimum Ph | 5.0-6.0 |
| Molecular Weight | ~57 kDa |
| Activity Concentration | ≥ 100,000 U/g |
| Solubility | Soluble in water |
| Storage Conditions | Store below 25°C in a cool dry place |
| Shelf Life | 12 months |
As an accredited Beta-Amylase factory, we enforce strict quality protocols—every batch undergoes rigorous testing to ensure consistent efficacy and safety standards.
| Packing | Beta-Amylase is packaged in sealed 25 kg fiber drums with inner polyethylene liner to prevent moisture absorption. |
| Container Loading (20′ FCL) | 20′ FCL: standard dry container, palletized bags/drums of Beta-Amylase, securely braced, protected from moisture, with proper labeling and ventilation. |
| Shipping | Beta-Amylase should ship refrigerated or frozen to preserve enzymatic activity. Use insulated containers with ice packs or dry ice, and expedited delivery. Avoid temperature fluctuations, prolonged exposure to heat, and moisture. Package securely to prevent leakage. Include clear labeling and documentation for biological/enzyme shipments. |
| Storage | Store Beta-Amylase in a tightly sealed, desiccated container at -20°C, protected from light and moisture. For solutions, aliquot before freezing to avoid repeated freeze-thaw cycles, which reduce activity. Under these conditions, the enzyme remains stable for extended periods. Always allow the product to equilibrate to the appropriate temperature before opening. |
| Shelf Life | Store refrigerated, sealed, and dry; shelf life typically 12 months. Avoid heat and humidity to retain activity. |
At a liquefaction DE of 8–12, a starch slurry of 30–35 g/100 g dry substance is cooled from jet cooking at 105–108°C to 58–60°C and adjusted to pH 5.0–5.5. Corn, tapioca, and wheat starch all enter the saccharification vessel after thermostable alpha-amylase has reduced viscosity and produced dextrins with an average degree of polymerisation below 12. Beta-amylase from barley malt or microbial fermentation is added as a liquid concentrate standardized to 700–750 BAU/g; typical starting doses range from 0.25 to 0.60 kg per metric ton dry solids. Pullulanase is co-dosed at 0.20 to 0.35 kg per metric ton dry solids to hydrolyse alpha-1,6 branch points, because beta-amylase alone stalls at residual branched dextrins. The saccharification vessel is a baffled steel tank with a top-entry agitator running at 20–30 rpm and an external cooling loop to hold temperature within ±1°C. Saccharification time ranges from 48 to 72 h. Maltose formation is monitored by HPAEC-PAD calibrated against ISO 17025 reference materials; DE is checked by reducing sugar titration according to ISO 5377:1981. Reversion product formation accelerates above pH 5.5, so pH is maintained with 0.5 M sodium carbonate dosed through a proportional pump.
The resulting hydrolysate contains 70–80 g maltose per 100 g carbohydrate, 1–2 g glucose per 100 g carbohydrate, and the balance as DP3+ dextrins. After enzyme inactivation at 100°C for 15 min, the syrup is clarified by rotary vacuum precoat filtration and demineralised through cation-anion ion exchange columns. The terminal product is a high-maltose syrup at 80–82 g/100 g dry substance, used in confectionery, brewing wort extenders, and fermentation media. Regulatory compliance for the enzyme preparation is governed by EU Regulation (EC) No 1332/2008 and the JECFA general specifications for food enzymes; the enzyme must be free of antimicrobial activity and mycotoxins.
| Substrate | Liquefaction DE | Beta-amylase dose (kg/t dry solids) | Pullulanase dose (kg/t dry solids) | Saccharification time (h) | Maltose end point (g/100 g carbohydrate) |
|---|---|---|---|---|---|
| Corn starch | 10–12 | 0.45–0.60 | 0.20–0.30 | 60–72 | 75–80 |
| Tapioca starch | 8–10 | 0.35–0.50 | 0.25–0.35 | 48–60 | 78–82 |
| Wheat starch | 10–12 | 0.50–0.70 | 0.25–0.35 | 60–72 | 70–76 |
The values in the table are representative starting ranges and require pilot confirmation for each substrate lot and enzyme batch.
The primary limiting factor is thermolability during the saccharification rest, not catalytic efficiency. In a lauter tun receiving 30–45% rice or corn adjuncts, the cereal side is jet cooked with thermostable alpha-amylase at 95–100°C before being combined with malt grist and cooled to 62–64°C. Exogenous beta-amylase is dosed into the combined mash at 0.05–0.15 L per metric ton of adjunct dry weight once pH stabilises at 5.3–5.5. The mash rest lasts 60–90 min. Native malt beta-amylase is assayed as diastatic power by ASBC Malt-3; process specifications commonly require a minimum of 60° DP for 100% malt and supplementary beta-amylase when adjunct exceeds 30% of grist. Fermentability limits are measured by apparent attenuation limit after forced fermentation; the target is ≥82% with a terminal maltose:glucose ratio above 2:1 in the wort. If the saccharification rest exceeds 65°C for more than 10 min, beta-amylase inactivation leaves limit dextrins and attenuates below specification.
Compliance in European brewing requires enzyme preparations to meet EU Regulation (EC) No 1332/2008; in the United States, TTB 27 CFR Part 25 formula approval may be required when enzyme is added outside standard brewing adjuncts. Recordkeeping of lot number, enzyme activity, and mash temperature is maintained in the brewhouse batch report. The terminal product is a fermentable wort that passes to pitched yeast with maltose as the principal carbon source, enabling reproducible ethanol and ester profiles in finished beer.
In lean doughs, beta-amylase activity from barley malt flour or wheat malt flour supplies maltose to Saccharomyces cerevisiae during the first 20–30 min of fermentation. Damaged starch content above 8% as measured by AACC 76-31.01 creates additional non-reducing ends, but beta-amylase acts only on gelatinised or mechanically degraded starch surfaces; the enzyme does not liquefy native starch granules. A typical substitution is 0.25–0.75 g barley malt flour per 100 g flour when flour diastatic activity is below 50 DP on a 14% moisture basis. The enzyme requires an aqueous dough phase at pH 4.5–5.5 and proofing temperature 32–35°C; activity is lost when crumb temperature exceeds 70°C during baking. The resulting maltose pool supports yeast CO2 production and crust colour via Maillard reactions. Excessive malt flour above 1% can create a sticky dough and low loaf volume, so farinograph absorption is adjusted by 0.5–1.0 percentage points. Terminal products are pan bread, rolls, and buns with controlled crumb texture and crust browning. Regulatory compliance in the EU follows EU Regulation (EC) No 1332/2008 for food enzymes; barley malt flour itself is a food ingredient and must comply with EU Regulation (EC) No 1881/2006 for mycotoxin maxima.
In industrial fermentation carbon feed, residual glucose above 1.5 g/100 mL triggers catabolite repression of regulated promoters such as lac, tac, and araBAD. Beta-amylase hydrolysis is therefore used to produce fermentation-grade maltose syrup with a low glucose fraction. Corn starch is liquefied at 105°C with thermostable alpha-amylase to a DE of 8–10, then cooled to 58–60°C and adjusted to pH 5.0–5.2. Beta-amylase is dosed at 0.30–0.55 kg per metric ton dry solids with pullulanase at 0.20–0.30 kg per metric ton dry solids. The 48–60 h saccharification is monitored by HPAEC-PAD; the endpoint is maltose >80 g/100 g carbohydrate and glucose <1.5 g/100 g carbohydrate. The liquid is refined by ion exchange and vacuum evaporation, then spray dried to maltose monohydrate with residual moisture 5.0–6.5 g/100 g and bulk density 0.55–0.70 kg/L. The terminal product is a fermentation carbon source for recombinant protein production, antibiotics manufacturing, and yeast propagation. Media preparation requires sterile filtration at 0.22 µm after reconstitution; the powder must be free of heavy metals above FCC limits and endotoxin below 0.5 EU/mg if used in injectable-grade intermediates.
If beta-amylase is added after jet cooking in a grain distilling mash, the hold time is set by the temperature drop from 105°C to 58°C. Whole corn, wheat, or rye slurry is liquefied with thermostable alpha-amylase at 105°C for 5–10 min; the mash then passes through a vacuum flash to 58–60°C. pH is adjusted to 5.0–5.2 with 0.5 M sulphuric acid before beta-amylase is metered at 0.3–0.6 L per metric ton of grain. The saccharification hold lasts 90–120 min in an insulated tank with low-shear agitation at 15–20 rpm. The hydrolysis product shifts from high-molecular dextrin to maltose, with the maltose:glucose ratio reaching 3:1 to 5:1. If the temperature exceeds 63°C for more than 10 min, beta-amylase inactivation leaves unfermentable limit dextrins and the subsequent yeast fermentation stalls below target final gravity. Terminal product is a fermentable wash for potable alcohol or neutral spirit distillation. In the United States, distilled spirits plants maintain ingredient and process records under 27 CFR Part 19; enzyme additions must be traceable by lot and volume. In the EU, potable alcohol production follows Regulation (EU) 2019/787 for spirit drinks, and enzyme preparations must comply with Regulation (EC) No 1332/2008.
Maintain pH below 5.5 in the beta-amylase saccharification vessel, because higher pH accelerates transglycosylation and reversion products such as maltotriose and panose. These products raise syrup viscosity and alter hard candy and coating behaviour. High-maltose syrup intended for confectionery is produced from corn starch with beta-amylase dosed at 0.30–0.50 kg per metric ton dry solids and pullulanase at 0.20–0.30 kg per metric ton dry solids; the saccharification endpoint is 70–80 g maltose per 100 g carbohydrate. After inactivation at 100°C for 15 min, the syrup is filtered, carbon-decolored to colour below 20 ICUMSA units, and ion-exchanged to conductivity below 50 µS/cm. The terminal product is evaporated to 80–85 g/100 g dry substance and either shipped as heavy syrup or blended into confectionery formulations. Viscosity is controlled by DE and oligosaccharide profile; confectionery manufacturers monitor viscosity by Brookfield rotational viscometer at 50°C and pH by direct electrode. EU compliance for the finished confectionery syrup follows Regulation (EC) No 1333/2008 on food additives only where relevant; the enzyme itself must comply with Regulation (EC) No 1332/2008.
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β-Amylase (EC 3.2.1.2, CAS 9000-91-3) is an exo-acting 4-α-D-glucan maltohydrolase that removes successive maltose units from the non-reducing ends of amylose, amylopectin, and related α-1,4-glucans. The enzyme belongs to glycoside hydrolase family GH14 and hydrolyses with inversion of the anomeric configuration, releasing β-maltose. Industrial product models are differentiated by source organism—barley, wheat, sweet potato, soybean, or microbial fermentation—and by delivery format, typically an amber liquid concentrate or a lyophilized powder. Commercial specifications list activity on a certificate of analysis using diastatic power (DP°) for brewing grades or defined enzyme activity units for food and starch processing grades. Plant-derived β-amylases are monomeric or tetrameric glycoproteins; the barley enzyme has a reported molecular mass near 59–60 kDa. Because the active site accommodates a defined number of glucosyl residues, hydrolysis is not complete on amylopectin. The enzyme stops 2–3 glucose residues from α-1,6 branch points, leaving β-limit dextrin as a co-product. Products described as β-amylase therefore vary by source, thermal tolerance, and unit definition, but share the same exo-acting maltogenic mechanism.
In starch conversion operations, β-amylase is not a liquefaction enzyme. It is added after gelatinization and partial hydrolysis, when soluble dextrins are present and the process temperature has been reduced to the enzyme’s operational window. Unconverted raw starch or ungelatinized granules reduce the concentration of accessible non-reducing ends and lower the observed reaction rate. The enzyme requires a free non-reducing chain end; very short maltodextrins are converted more slowly because the substrate cannot occupy the multiple subsites of the active site cleft. This property influences dosage calculations in high-maltose syrup production and mash tun supplementation, where residual starch carried into the kettle reduces real extract and final attenuation.
The operational distinction among amylolytic enzymes is defined by attack mode, linkage specificity, and product distribution. α-Amylase (EC 3.2.1.1) is an endo-acting enzyme that hydrolyses internal α-1,4 linkages, rapidly reducing viscosity and generating branched and linear dextrins. It does not release maltose selectively and is therefore used for liquefaction rather than sugar profiling. β-Amylase is exo-acting and releases maltose from the chain terminus, but cannot bypass α-1,6 branch points. Glucoamylase (EC 3.2.1.3) is also exo-acting but releases β-D-glucose and hydrolyses α-1,6 linkages at a slower rate, making it suitable for high-glucose and fermentation ethanol operations. Pullulanase (EC 3.2.1.41) and isoamylase (EC 3.2.1.68) cleave α-1,6 branch linkages and are therefore used in combination with β-amylase to improve maltose yield by removing the structural barrier that causes β-limit dextrin accumulation.
In a mixed α-amylase/β-amylase system, α-amylase produces new non-reducing ends for β-amylase attack, while β-amylase consumes short linear segments and accumulates maltose. In a β-amylase/glucoamylase blend, glucose formation rises at the expense of maltose because glucoamylase hydrolyses maltose and higher oligosaccharides. Control of blend ratio and dosing sequence therefore determines the final carbohydrate spectrum.
| Enzyme | EC code | Mode | Linkage cleaved | Principal product | Branch activity |
|---|---|---|---|---|---|
| α-Amylase | 3.2.1.1 | Endo | α-1,4 | Dextrins and oligosaccharides | None |
| β-Amylase | 3.2.1.2 | Exo | α-1,4 | Maltose | None |
| Glucoamylase | 3.2.1.3 | Exo | α-1,4 and α-1,6 | Glucose | Slow |
| Pullulanase | 3.2.1.41 | Endo | α-1,6 | Linear chains | Specific |
Commercial β-amylase is sold under source-specific trade names, but the technical specification is organized around activity, pH, temperature response, physical format, and microbiological quality. Liquid brewing grades are commonly specified by diastatic power using ASBC Malt-7A or EBC 4.11, in which dextrinized starch is incubated with the sample and reducing sugars are titrated iodometrically. Food processing grades may use soluble starch or blocked p-nitrophenyl maltooligosaccharide substrates, with one unit defined as the release of 1 μmol of product per minute under stated temperature and pH. The certificate of analysis must state the unit basis, because unit definitions are not interchangeable across suppliers or source organisms. Typical liquid preparations have pH 4.8–5.8 and density 1.05–1.15 g/mL; powder preparations are hygroscopic and should be handled at less than 40% relative humidity. Liquid formulations may contain stabilizers and preservatives; powder preparations may be standardized with dextrins or maltodextrins to a defined activity.
Barley β-amylase has optimum pH 4.6–5.6 and optimum temperature 50–55 °C in dilute buffered starch assays. Sweet potato β-amylase is reported to retain activity at 60–65 °C, and some microbial β-amylases from Bacillus species are active at 60–70 °C. These optima shift with substrate concentration, pH, soluble solids, and calcium or salt level; process validation is required before changing source organism. Published data for each specific configuration is limited at high dry solids above 30% DS, where viscosity and product inhibition change apparent kinetics.
Within the β-amylase product class, source organism creates measurable differences. Plant-derived β-amylases from barley and sweet potato are used where maltogenic character is desirable, but they may be less thermostable and more sensitive to heavy-metal inhibition. Microbial β-amylases from Bacillus strains are selected for higher reaction temperatures and lower colour formation in starch hydrolysates. The selection of a specific product model therefore depends on dry solids, target reaction time, and final syrup colour specification. A liquid barley β-amylase product is not interchangeable with a microbial powder product solely on the basis of activity units; thermal deactivation rate and viscosity response must be checked under process conditions.
| Standard or regulation | Scope | Acceptance criterion |
|---|---|---|
| JECFA enzyme monograph | Identity and purity of food enzyme preparations | Complies with microbial limits and permitted processing aids |
| FCC enzyme specifications | Activity and food chemical codex status | Declared activity within lot-specific range |
| ASBC Malt-7A | Diastatic power in brewing products | Method-specific DP result |
| EBC 4.11 | Diastatic power in European malt analysis | Method-specific DP result |
| Regulation EU 1332/2008 | Authorisation of food enzymes in the EU | Union list status applicable to the specific source |
| ISO 22000 | Food safety management at manufacturing site | Certified system for enzyme production |
In brewing, β-amylase is endogenous to malt, where it determines the fermentable extract available to yeast. Malt specifications for all-malt lager grists often require diastatic power above 110 °ASBC on a dry basis. When raw barley, corn, or rice adjuncts exceed 30% of the grist, low malt DP becomes process-limiting because unconverted starch carries into the mash filter and kettle, reducing extract yield and increasing haze risk. Exogenous β-amylase is added to the mash at 55–65 °C and pH 5.2–5.6, with dose determined by the difference between target and measured malt DP. In production-scale brewhouses using 200–500 hL mashing vessels, enzyme is diluted in cold water and dosed through a separate line to avoid local protein precipitation. Fermentability is monitored by wort attenuation limit or real degree of fermentation, not by enzyme activity alone.
In high-maltose syrup manufacture, β-amylase is combined with a debranching enzyme after liquefaction. Thermostable α-amylase gelatinizes starch at 105–110 °C, then the stream is cooled to 55–60 °C and adjusted to pH 5.0–5.5. β-Amylase and pullulanase are dosed into the saccharification vessel and held for 24–72 h. The resulting syrup commonly contains 50–60% maltose, less than 5% glucose, and 15–25% maltotriose, but the actual profile depends on dextrose equivalent, calcium, dry solids, and debranching efficiency. Product specification for high-maltose syrup requires HPLC verification of the saccharide spectrum, typically using AOAC 977.08 or equivalent carbohydrate column methods. Excess β-amylase dosage does not generate glucose; unconverted β-limit dextrins remain and may require further treatment with glucoamylase if a fully fermentable stream is required.
In baking applications, β-amylase is used at low dosages to increase fermentable maltose for yeast and to alter crumb structure in lean doughs. The addition level is limited because excessive maltose increases Maillard browning and may produce sticky crumb after baking. Process validation through farinograph and extensigraph testing is required, using methods such as AACC 54-21 and AACC 54-10. Published data for specific dough systems is limited; production-scale validation under actual flour specifications is necessary.
Analytical use of β-amylase includes preparing β-limit dextrins for starch fine-structure determination. Treatment conditions follow the specific assay protocol, usually at pH 4.8–5.5 and 40–55 °C, until no further reducing sugar is released. The β-limit value is then determined by iodine binding, size-exclusion chromatography, or reducing-sugar measurement.
Thermal stability is a primary operational boundary. Barley β-amylase denatures rapidly above 60–65 °C; a 5 °C overshoot in a diluted enzyme line can reduce activity by 50% within 15–30 min. Microbial β-amylases tolerate higher temperatures, but the same principle applies: the enzyme should be added only after the process stream is below the supplier’s declared inactivation threshold. Holding β-amylase in a hot dosing line without flow creates dead-volume inactivation and lot-to-lot variation in downstream sugar composition.
pH below 4.0 reduces catalytic activity through protonation of active-site residues, while pH above 7.0 accelerates denaturation for many plant-derived preparations. The enzyme is incompatible with strong oxidizing agents such as peracetic acid, chlorine dioxide, and hypochlorite sanitizer residues. Dosing lines should be flushed with potable water after clean-in-place cycles and verified with test strips or ATP swabs before enzyme introduction. Heavy-metal ions, particularly Cu²⁺ and Hg²⁺, inhibit plant β-amylase; brass or copper fittings should be avoided. Avoid premixing β-amylase with alkaline cleaning solutions, cationic flocculants, or concentrated salts that precipitate protein.
Storage recommendations for liquid β-amylase are 4–8 °C, with powder storage at ≤25 °C and relative humidity below 40%. Freezing of liquid formulations can cause protein aggregation and activity loss. Opened containers should be used within the supplier’s stated post-opening window because preservative systems are designed for sealed conditions. Published stability data for dilute enzyme solutions in high-solids syrups are limited; pilot-scale inactivation studies under actual conversion conditions are recommended before changing source organism or formulation.
