Optimizing red sorghum malt quality when Bacillus subtilis is used during steeping to control mould growth

Previous work has shown that Bacillus subtilis‐S499‐based biocontrol treatments applied without aeration at the steeping stage of red sorghum malting offer good mould reduction, but yield malts with low levels of key hydrolytic enzymes. Thus we attempted to raise these levels by aerating the steeping liquor, varying the steeping time (from 8 to 40 h) and temperature (from 25 to 35 °C), and combining a biocontrol treatment with prior steeping in 0.2% NaOH. Aeration proved particularly important whenever B. subtilis cells were present in the steep liquor. The optimal temperatures for α‐ and β‐amylase were 30 and 25 °C, respectively. By increasing the steeping time, it was possible to improve the α‐amylase activity, but the β‐amylase activity peaked sharply between 16 and 20 h, depending on the steeping medium. A good compromise was steeping in a biocontrol medium for 14–16 h at 30 °C. Combination steeping treatments (0.2% NaOH for 8 h followed by biocontrol for 8 h) yielded malts of a quality approaching that afforded by dilute alkaline treatment. Copyright © 2012 The Institute of Brewing & Distilling     

Malting of Sorghum: Further Studies on Factors influencing α‐Amylase Activity

The effects of steep regime, nature of alkaline steeping agent, and kilning condition on α‐amylase development were studied for four Nigerian sorghum cultivars. Malt α‐amylase activity was highly significantly (p<.001) influenced by all the four factors as well as their various assortments of interaction. Generally malts from the Local Red (LR) variety produced the highest a‐amylase values, followed by those of SK 5912, Local White and KSV 8 in the above sequence. The presence/absence of air‐rest processes in steep regimes was a significant factor (p<.001) influencing malt α‐amylase response to final warm steeping as well as to the other factors under study. Similarly, the nature of the steeping agent was a very significant determinant of malt α‐amylase response to kilning condition and regime of steeping. Of significant interest was the observation that Ca (OH)₂ steeps enhanced malt α‐amylase activity at the higher temperature of kilning. The significantly lower α‐amylase values given under similar conditions by the other alkaline liquors suggest a possible increase in malt thermostability due to steeping in Ca (OH)₂. Additionally, the fact that the extent of enhancement of malt α‐amylase activity by Ca (OH)₂, at 50°C Kiln temperature, was regime‐dependent, suggests that the latter was an important modulator of sorghum germination physiology.     

Sorghum β‐Amylase Production: Relationship With Grain Cultivar,Steep Regime,Steep Liquor Composition and Kilning Temperature

Steep regime, nature of alkaline liquor, and kilning conditions were studied for their effects on sorghum malt β‐amylase development in four Nigerian sorghum cultivars. Malt β‐amylase activity was markedly (p < .001) influenced by all the four factors as well as their various interactions. Overall, malts from KSV 8 variety were the most β‐amylolytic followed in sequence by those from Local Red (LR), SK 5912, and Local white (LW) respectively. The presence or absence of air rests in steep regimes was a significant (p < .001) determinant of sorghum β‐amylase response to final warm steeping, steep liquor and kilning condition. The nature of the alkaline steep liquor was also a major determinant of the pattern of malt β‐amylase response to the kilning condition. Steeping in Ca(OH)₂ enhanced malt β‐amylase activity at the higher temperature of kilning, while KOH produced the opposite effect. Ca(OH)₂ enhancement of β‐amylase development, at a kilning temperature of 50°C, was variety‐dependent suggesting that different sorghum cultivars may employ different biosynthesis models for β‐amylase synthesis. The regime‐dependence of β‐amylase response to kiln temperature suggests that this was an important modulator of sorghum germination physiology.     

Characterization of an α‐amylase from sorghum (Sorghum bicolor) obtained under optimized conditions

Sorghum malt α‐amylase can compete with bacterial α‐amylase in industrial applications, if sufficiently stable and produced in a large enough quantity. Conditions for maximal α‐amylase production in sorghum malt and the physico‐chemical properties of the α‐amylase so produced are reported in this study. Sorghum grains were steeped in buffers with varying pH (4.0–8.0) for 24 h, at room temperature, and germinated for another 48 h to obtain the green malt. The buffer that induced the highest quantity of α‐amylase was chosen as the optimal pH and served as the medium for further steeping experiments conducted at different temperatures (10, 20, 30, 40, 50 and 60°C). The α‐amylase activity in the extract was determined in order to obtain the optimum temperature for α‐amylase induction at this particular pH. For the purpose of comparison, the α‐amylase produced at the optimum pH and temperature was purified to apparent homogeneity by a combination of ion‐exchange and size‐exclusion chromatography, and further characterized. Eight‐fold higher α‐amylase activity was induced in pH 6.5 buffer at 20°C compared with water, the traditional steeping medium. The K_m and V_max were estimated to be 1.092 ± 0.05 mg mL^?1 and 3516 ± 1.981 units min^?1, respectively. The activation energy of the purified amylase for starch hydrolysis was 6.2 kcal K^?1 mol^?1. Chlorides of calcium and manganese served as good activators, whereas CuSO₄ inhibited the enzyme with a 42% loss in activity at 312 mm salt concentration. Copyright © 2012 The Institute of Brewing & Distilling     

Effect of cold shock on the enhancement of β‐amylase activity during malting and malt processability for a red sorghum intended for brewing use

The low β‐amylase activity of sorghum malt is a major concern when malts are intended for use in brewing. Several studies have shown that the germination temperature plays an important role in β‐amylase synthesis. In this study, the cold shock treatment was envisioned as a means of improving β‐amylase synthesis during red sorghum malting. The results show that, when a high‐frequency decrease in the germination temperature is used, the obtained malt exhibits a significantly increased β‐amylase activity. This study shows that this increase is not sufficient to consider cold shock as a means of improving β‐amylase activity for red sorghum brewing use, as the processabilty of the malts is unsatisfactory. Copyright © 2015 The Institute of Brewing & Distilling     

The influence of steep regime and germination period on the malting properties of some hull‐less barley lines

Following field trials in 2008 and 2010, six lines, derived by mutation in the hull‐less barley cultivar Penthouse, were selected to provide a range in grain β‐glucan content. These lines, along with Penthouse and the hulled, malting variety Optic, were then malted, using four different steeping regimes, with samples kilned after 3, 4 and 5 days of germination. The longest steep regime provided samples with optimum modification after 5 days of germination. Samples from the other steep regimes were under‐modified to varying degrees. In particular, the steeps with a single immersion gave poorly modified samples with low extracts and alcohol yields. One line, with low grain β‐glucan, gave higher alcohol yields than either Penthouse or Optic, following a regime comprising two short immersions and a single air‐rest, but there was no clear association, within the lines, between β‐glucan content and malting properties. Copyright © 2012 The Institute of Brewing & Distilling     

Control of Superoxide Dismutase Activity during Malting Using Plackett‐Burman and Box‐Behnken Experimental Design and Its Effect on Reducing Power of Wort

The Plackett‐Burman multifactorial design was employed to screen the important malting parameters for superoxide dismutase (SOD) in final malt of Ganpi‐3. The eight factors screened for SOD were steeping temperature, steeping time, peroxide hydrogen concentration in steeping water, germination temperature, germination time, withering temperature, drying temperature and kilning temperature. Variance analysis showed that steeping time, germination temperature and kilning temperature were significant for SOD activity. Box‐Behnken experimental design was further used to optimize the levels of the above three factors. By response surface methodology and canonical analysis, the optimal malting factors for higher SOD activity in final malt were: steeping time 42.2 h, germination temperature 16.9°C and kilning temperature 82.9°C. Under these conditions, the model predicted a SOD activity of 2234 U/g of dry weight malt. Verification of the optimization showed that a SOD activity of 2220 U/g was observed under optimal conditions. It showed that the experimental data could be reliably predicted by the polynomial model. Besides Ganpi‐3, three other barley varieties including Ganpi‐4, Ken‐2 and Hamelin were malted under optimal and common conditions under laboratory conditions. To some extent, SOD activities were higher in malts from the optimal malting process than those from the common malting process. Especially, SOD activities in Ganpi‐3 and Hamelin increased by 18.8% and 15.3%, respectively. Furthermore, twenty‐nine samples of malts, including eleven imported malts and eighteen domestic malts, were used. Relationships between SOD activity in malt and the reducing power of wort were examined. There was significant correlation between SOD activity and the reducing power of wort (R² = 0.8069).     

α‐(1,4)‐Amylase,but not α‐ and β‐(1,3)‐glucanases,may be responsible for the impaired growth and morphogenesis of Paracoccidioides brasiliensis induced by N‐glycosylation inhibition

The cell wall of Paracoccidioides brasiliensis, which consists of a network of polysaccharides and glycoproteins, is essential for fungal pathogenesis. We have previously reported that N‐glycosylation of proteins such as N‐acetyl‐β‐d ‐glucosaminidase is required for the growth and morphogenesis of P. brasiliensis. In the present study, we investigated the influence of tunycamicin (TM)‐mediated inhibition of N‐linked glycosylation on α‐ and β‐(1,3)‐glucanases and on α‐(1,4)‐amylase in P. brasiliensis yeast and mycelium cells. The addition of 15 µg/ml TM to the fungal cultures did not interfere with either α‐ or β‐(1,3)‐glucanase production and secretion. Moreover, incubation with TM did not alter α‐ and β‐(1,3)‐glucanase activity in yeast and mycelium cell extracts. In contrast, α‐(1,4)‐amylase activity was significantly reduced in underglycosylated yeast and mycelium extracts after exposure to TM. In spite of its importance for fungal growth and morphogenesis, N‐glycosylation was not required for glucanase activities. This is surprising because these activities are directed to wall components that are crucial for fungal morphogenesis. On the other hand, N‐glycans were essential for α‐(1,4)‐amylase activity involved in the production of malto‐oligosaccharides that act as primer molecules for the biosynthesis of α‐(1,3)‐glucan. Our results suggest that reduced fungal α‐(1,4)‐amylase activity affects cell wall composition and may account for the impaired growth of underglycosylated yeast and mycelium cells. © 2013 The Authors. Yeast published by John Wiley & Sons Ltd.     

The Influences of Steeping Duration and Temperature on the α‐ and β‐Amylase Activities of Six Thai Rice Malt Cultivars (Oryza sativa L. Indica)

A preliminary study of malting conditions for six Thai rice cultivars was conducted. Three non‐glutinous rice cultivars (KDML105, PT60, and WR) and three glutinous rice cultivars (SPT, RD6, and KND) were selected. The steeping durations (24, 48, and 72 h) and temperatures (20, 25 and 30°C) were investigated for their effect on α‐ and β‐amylase, the key enzymes for malt quality evaluation. During steeping, the production of both enzymes was lower than at the germination process. The longer the steeping duration, the lower the maximum β‐amylase activity obtained. The contradictory effect was observed for α‐amylase activity, near the end of the germination time. Additionally, temperature influenced the water absorption content as well as the amylolytic enzyme activity. Particularly at 30°C, the maximum β‐amylase activity (6.7 unit/mg protein) was found in KND malt steeped for 24 h, and maximum α‐amylase activity (20 unit/mg protein) was found in PT60 malt steeped for 72 h. The amount of enzyme production depended on the variety rather than the amylose content in the rice. The optimal condition for malting rice regarding β‐amylase activity and α‐amylase activity was analyzed at 30°C, with steeping for 24 h and germination for 4–5 days.     

Effects of germination on the activities of amylases and phenolic enzymes in sorghum varieties grouped according to food end‐use properties

Fifty sorghum varieties were screened to determine the effects of germination on levels of starch, α‐amylase, β‐amylase, phenylalanine ammonia lyase (PAL), peroxidase (POX) and polyphenol oxidase (PPO). Germination decreased starch content, with amylose being more degraded than amylopectin. In germinated grain, α‐amylase activity increased several‐fold in all varieties, whereas β‐amylase activity did not increase uniformly and even decreased in some varieties. Activity of the key enzyme in phenolic biosynthesis, PAL, was detected in only half of the varieties before germination but in all of them after germination. PPO was not activated in germinated sorghum grains, whereas POX activity increased up to tenfold in some varieties. Zymography revealed that germination induced de novo synthesis of several POX isoenzymes, among which an anionic POX isoenzyme (pI 3.1) was ubiquitously present. Amylase and phenolic enzyme activities could be correlated with grain and plant agronomic characteristics. The use of sorghum varieties for local dishes such as ‘tô’, ‘dolo’, couscous and thin porridge could be correlated with amylase and phenolic enzyme activities and the contents of their substrates. The biochemical constituents determined are useful markers for selection of varieties for food utilisation with special emphasis on infant porridges. Copyright © 2006 Society of Chemical Industry     

Phytosterols in banana (Musa spp.) flower inhibit α‐glucosidase and α‐amylase hydrolysations and glycation reaction

Three phytosterols were isolated from Musa spp. flowers for evaluating their capabilities in inhibiting glucosidase and amylase activities and glycation of protein and sugar. The three phytosterols were identified as β‐sitosterol (PS1), 31‐norcyclolaudenone (PS2) and (24R)‐4α, 14α, 4‐trimethyl‐5α‐cholesta‐8, 25(27)‐dien‐3β‐ol (PS3). IC₅₀ values (the concentration of inhibiting 50% of enzyme activity) of PS1, PS2 and PS3 against α‐glucosidase were 283.67, 11.33 and 43.10 μg mL^?1, respectively. For inhibition of α‐amylase, the IC₅₀ values of PS1, PS2 and PS3 were 52.55, 76.25 and 532.02 μg mL^?1, respectively. PS1 was an uncompetitive inhibitor against α‐amylase with K_m at 5.51 μg mL^?1, while PS2 and PS3 exhibited a mixed‐type inhibition with K_m at 52.36 and 2.49 μg mL^?1, respectively. PS1 and PS2 also significantly inhibited the formation of advanced glycation end products (AGEs) in a BSA–fructose model. The results suggest that banana flower could possess the capability in prevention of the diseases associated with abnormal blood sugar and AGEs levels, such as diabetes.     

Variety and germination time effect on total β‐glucan,water‐insoluble β‐glucan,water‐soluble β‐glucan components and β‐glucanase levels in improved sorghum varieties SK5912, KSV8 and ICSV400 before and after malting and their relationships to wort viscosity

The effects of variety and germination time on β‐glucan components – total β‐glucan (TBG), water insoluble β‐glucan (WIBG) and water soluble β‐glucan (WSBG) and β‐glucanase (BG) levels – before and after malting in improved sorghum varieties SK5912, KSV8 and ICSV400 and their relationships to wort specific viscosity (SV) were studied. This study was part of efforts to aid local malting and brewing industries in the application of sorghum varieties that are abundantly available to reduce costs. At the fifth day of germination, variety ICSV400 had the lowest TBG, WIBG and WSBG levels in its raw and malt samples. Variety SK5912 had the highest TBG, WIBG and WSBG levels in its raw samples, while variety KSV8 had the highest levels of TBG, WIBG and WSBG in its malt samples. Similarly, variety ICSV400 malts developed the highest BG levels, while the KSV8 malts gave the lowest level. The effect of variety, germination time and variety × germination time interaction was significant (p < 0.05) on the TBG, WIBG and BG levels and was not significant on the WSBG levels. Weak and significant correlation of TBG levels with SV (0.25, p < 0.05 for SK5912; 0.24, p < 0.05 for KSV8; and 0.31, p < 0.05 for ICSV400) was observed in all the samples, suggesting that the low β‐glucan levels may not be primarily and solely responsible for any viscosity impediments associated with sorghum worts during run‐off. With improvement in the effective utilization of sorghum, ICSV400 appeared the most suitable variety for malting and brewing in Nigeria.Copyright © 2016 The Institute of Brewing & Distilling     

Laboratory Wet‐Milling of Grain Sorghum With Abbreviated Steeping to Give Two Products

Short‐time and no‐time steeping were used in the wet‐milling of grain sorghum to give two products, starch in over 78% recovery (starch basis) plus the remaining grain solids. In the wet‐milling process 1.5 parts of fresh water were used per part of grain to compensate for drying and transfer losses. Starting with 100 g (dry solids) of commercial No.2 grain sorghum, steep time (1—3 h), steep temperature (25—60 °C), and coarse‐grinding speed (7, 500—12, 500 rpm with tip speed of 90.4—150.7 km/h) were varied in a model study; starch recovery, starch lightness (L*), and damaged starch were the responses. Grain sorghum was steeped with twice its weight of process water containing 0.2% sulfur dioxide and the steeped kernels were added to an equal volume of process water and the mixture was ground for 6 min in a Waring blender with blunt blades (d = 3.2 cm). The course‐ground material was sieved (opening 1190 μm) to collect the bran/germ, and the throughs were allowed to stand. The sedimented phase was finely ground by one pass through a plate mill, and the fine fiber removed by sieving (opening 73 μm). The slurry was adjusted to a specific gravity of 1.04, and the starch was separated from the protein fraction on a starch table. The protein fraction was combined with the steep‐liquor concentrate (54% solids) plus the bran/fiber and the fine‐fiber fractions to give the co‐product, which contained 70% moisture (wet basis, wb) and 27% protein (dry basis, db). In the surface response study, recovery of starch ranged from 57 to 89%, starch protein content from 0.4 to 0.5%, and lightness (L*) from 90 to 93. Damaged starch content was constant at around 0.4%. Commercial grain sorghum gave the highest starch recovery (90%) after steeping 2 h at 55 °C and coarse‐grinding at 10, 500 rpm; whereas a food‐grade, a white and a red sorghum gave 85, 84, and 80% recoveries, respectively. The four starches had lightness (L*) values of 93—94 and damaged starch contents of 0.4—0.6%. When the commercial grain sorghum was wet‐ground without steeping in 2 parts of water containing 0.2% sulfur dioxide, a 78% recovery of starch was obtained with L* 93.7 and starch damage 0.5%.     

Bacillus subtilis HJ18‐4 from Traditional Fermented Soybean Food Inhibits Bacillus cereus Growth and Toxin‐Related Genes

Bacillus subtilis HJ18‐4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad‐spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18‐4. Expression of B. cereus toxin–related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18‐4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18‐4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18‐4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus.     

Isolation of a Recombinant Bacillus sp. TS‐23 α‐Amylase by Adsorption‐Elution on Raw Starch

A Bacillus sp. TS‐23 α‐amylase produced by recombinant Escherichia coli was adsorbed onto raw starch and the adsorbed enzyme was eluted with maltose or maltodextrin in 50 mM Tris/HCl buffer (pH 8.5). The adsorption‐elution procedure resulted in a yield of 53% α‐amylase activity and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS/PAGE) analysis showed that the eluted α‐amylase had a molecular mass of approximately 64 kDa. Raw starch could be used repeatedly in the adsorption‐ elution cycle with good reproducibility. Scanning electron microscopy of the isolated corn starch exhibited a smooth appearance of the granules before adsorption and only a small change in appearance after three adsorption‐elution cycles. These results suggest that the raw starch adsorption‐elution technique has a great potential in the isolation of Bacillus sp. TS‐23 α‐amylase from the culture broth of recombinant E. coli.     

The Effect of Steeping Time on the Final Malt Quality of Buckwheat

To determine the effect of steeping time on final buckwheat malt quality, buckwheat was steeped for three different times resulting in three different out‐of‐steep moisture contents: 7 h steeping (35%), 13 h steeping (40%) and 80 h steeping (45%). An increased steeping time increased malting losses, total beta‐amylase activity and Kolbach index. On the contrary total nitrogen, friability and viscosity of consequent congress worts were decreased. A maximum alpha‐amylase activity was found in buckwheat malted with an out‐of‐steep moisture content of 45%. Beta‐amylase existed in a soluble and latent form in buckwheat. The latent form was solubilised during malting. In addition extra beta‐amylase was produced. In general the optimum out‐of‐steep moisture content for buckwheat is between 35 to 40%, which is a compromise between attaining the desired malt quality and minimising malting loss.     

Purification and enzymatic identification of an acid stable and thermostable α‐amylase from Rhizopus microsporus

Rhizopus microsporus, recently isolated from a solid culture of Heng‐Shui Lao‐Bai‐Gan (HSLBG, a famous distilled liquor in Northern China) was found to produce a novel extracellular acid stable and thermostable α‐amylase. This fungal α‐amylase was purified using ammonium precipitation, Sephadex G‐25 desalination and DEAE‐52 cellulose chromatography. Its molecular weight was estimated to be 75 kDa by SDS–PAGE. The optimum pH and temperature of this enzyme was pH 5.0 and 70°C respectively. Thermostability and kinetic analysis through the Arrhenius and Michaelis–Menten equations revealed that this enzyme showed an exceptional activity at low pH and high temperature. A combination of this thermostability and acid stability could be a valuable trait for the efficient hydrolysis of amylose to glucose in large‐scale biotechnology applications. Copyright © 2012 The Institute of Brewing & Distilling     

The Use of Response Surface Methodology to Optimise Malting Conditions of Proso Millet (Panicum miliaceum L.) as a Raw Material for Gluten‐Free Foods

Response surface methodology was used to investigate the influence of the three malting parameters, i.e. degree of steeping, germination time, and temperature on the quality of proso millet malt. Each predictor variable was tested at three levels. Germination times varied from 5, 6, and 7 days, degrees of steeping were 44, 48, and 52%, and germination temperatures were 16, 20, and 24°C. A set kilning temperature of 65°C was used for all malts. A series of malt quality parameters were investigated including extract, apparent attenuation limit, gelatinisation temperature, α‐amylase activity, β‐amylase activity, limit dextrinase activity, Kolbach index, α‐amino nitrogen, viscosity, and colour. The optimal malting program was achieved after the fifth day of germination, 44% degree of steeping, and a 22°C steeping and germination temperature. The obtained values for the amylolytic and cytolytic attributes were 64.8% extract, 1.383 mPa × s viscosity, 76.0% apparent attenuation limit, 111 U/g α‐amylase activity, and 102 U/g β‐amylase activity.     

Changes in enzyme activities and aflatoxin elaboration in sorghum genotypes following Aspergillus parasiticus infestation

Sorghum is a relatively poor substrate for aflatoxin production compared with high‐risk agricultural commodities like maize and groundnut, even though it is susceptible to fungal attack. Fungal infestation of sorghum results in a varied biochemical composition of the deteriorated grain. In this study, six sorghum genotypes (red—AON 486, IS 620; yellow—LPJ, IS 17 779; white—SPV 86, SPV 462) were inoculated with a toxigenic strain of Aspergillus parasiticus (NRRL 2999) in order to evaluate the changes in the activities of various hydrolytic enzymes (α‐ and β‐amylases, protease and lipase) in comparison with those in uninfected grains. Enzyme activities were measured at different times after fungal infestation, and the enzymatic activities were correlated with the aflatoxin production. Alpha‐amylase activity was observed to be greater than β‐amylase activity in all six genotypes under both healthy and infected conditions. The increase in α‐amylase activity during the period of infection was higher in white genotypes than in red sorghum genotypes. Alpha‐amylase activity in all the genotypes increased up to day 6 after fungal infection, but was significantly lower in infected grains than in healthy grains. The variability in the basal enzyme activities among the six sorghum genotypes was quite high compared with the amount of induction of each specific enzyme due to infection and germination. Higher protease activity was observed in the infected grains than in healthy grains. The enzyme activities in high tannin red genotypes were less than those in yellow and white genotypes. The α‐ and β‐amylase activities were positively correlated (r = 0.406 and 0.436; P < 0.05) to aflatoxin production. Inherent lipase activity was highest (on day 0) in AON 486, SPV 462 and SPV 86, as compared with the activity in infected grains. The total aflatoxins produced (quantified by TLC‐fluorodensitometry) were lower in red genotypes than in yellow and white genotypes, suggesting that red genotypes were least susceptible to aflatoxin elaboration among the various genotypes tested. All four aflatoxins, (B₁, B₂, G₁ and G₂) were present in five genotypes (IS 620, LPJ, IS 17 779, SPV 86 and SPV 462) at all the stages of infection, but, aflatoxin could not be detected in the red genotype AON 486 on day 3 after infection. White genotypes SPV 86 and SPV 462) showed maximal aflatoxin (total) production on day 6 after infection. © 2000 Society of Chemical Industry     

Inhibitory potential of phenylpropanoid glycosides from Ligustrum purpurascens Kudingcha against α‐glucosidase and α‐amylase in vitro

The leaves of Ligustrum purpurascens are used in a Chinese traditional tea called small‐leaved kudingcha, which is rich in phenylpropanoid glycosides (PPGs) and has many beneficial properties. Two critical exoacting glycoside hydrolase enzymes (glucosidases) involved in carbohydrate digestion are α‐glucosidase and α‐amylase. We investigated the properties of PPGs from L. purpurascens for inhibiting α‐amylase and α‐glucosidase activity in vitro and found IC₅₀ values of 1.02 and 0.73 mg mL^?1, respectively. The patterns of inhibiting both α‐amylase and α‐glucosidase were mixed‐inhibition type. Multispectroscopy and molecular docking studies indicated that the interaction between PPGs and α‐amylase and α‐glucosidase altered the conformation of enzymes, with binding at the site close to the active site of enzymes resulting in changed enzyme activity. Our studies may help in the further health use of small‐leaved kudingcha.