Amylase Synthesis Research Articles

Characterization of Plant Growth Promoting Rhizobacteria and Their Benefits on Soybean Growth

Background: Soybean is the most important legume in the world and its seeds contains 40% protein and 20% oil. In recent years, more attention has been paid to the use of plant growth-promoting (PGP) rhizobacteria as a biofertilizer alternative to chemical fertilizers, which may pose risks to the environment. The rhizobia inoculation of soybean is a sustainable practice to induce atmospheric nitrogen fixation and subsequently improve crop productivity and soil fertility. Methods: In this study, 19 indigenous rhizobia isolated from soybean were tested in vitro for their plant growth-promoting properties (PGPR), while genotypic characterization included sequencing of the 16S rRNA gene. Result: Morphological characterisation has shown that all of the isolates were rod-shaped gram negative bacteria. The sequencing has shown that 12 out of 19 isolates belong to the genus Pseudomonas, four of which belong to P. fluorescens species. Isolates which belong to the Pseudomonas genus have shown the highest ability of indole-3-acetic acid synthesis, phosphorous solubilization and along with isolates SGN6, SGN7 and SGS4 (Sphingomonas sanguinis) potassium solubilisation too. All the isolates from Pseudomonas genus as well as those belonging to B. japonicum species have shown the protease synthesis abilities while amylase synthesis abilities was observed only in the isolate SAK2 (P. chlororaphis). The most efficient strains in in vitro biological nitrogen fixation assay belong to Pseudomonas and Rhizobium genera. Most of the isolates were positive to the organic acid production while all of them have shown the ability of the exopolysaccharide production.

A key component Rxt3 in the Rpd3L histone deacetylase complex regulates development, stress tolerance, amylase production and kojic acid synthesis in Aspergillus oryzae.

Rpd3L is a highly conserved histone deacetylase complex in eukaryotic cells and participates in various cellular processes. However, the roles of the Rpd3L component in filamentous fungi remain to be delineated ultimately. In this study, we constructed two knockout mutants of Rpd3L's Rxt3 subunit and characterized their biological functions in A. oryzae. Phenotypic analysis showed that AoRxt3 played a positive role in hyphal growth and conidia formation. Deletion of Aorxt3 resulted in augmented tolerance to multiple stresses, including cell wall stress, cell membrane stress, endoplasmic reticulum stress, osmotic stress and oxidative stress. Noteworthily, we found that Aorxt3-deleting mutants showed a higher kojic acid production than the control strain. However, the loss of Aorxt3 led to a significant decrease in amylase synthesis. Our findings lay the foundation for further exploring the role of other Rpd3L subunits and provide a new strategy to improve kojic acid production in A. oryzae.

Production of Amylase Enzyme by Aspergillus and Fusarium Species using Sugar Cane Bagasse

Study’s Novelty/Excerpt This study presents insights into the cost-effective production of amylase enzymes using sugar cane bagasse as a substrate, showcasing a significant advancement over traditional synthetic starch substrates. By optimizing production parameters such as incubation time, substrate and inoculum concentrations, pH, and temperature, the research demonstrates the superior amylase yield of Aspergillus niger, Aspergillus flavus, and Fusarium spp. under specific conditions. This work not only highlights the potential of using waste materials for industrial enzyme production but also emphasizes the need for further exploration of the isolates' identities and the industrial applications of the extracted enzymes. Full Abstract Amylases are hydrolytic enzymes produced by some microorganisms and utilized in the hydrolysis of starch. This study aimed to determine the ability of certain fungal isolates isolated from wastes to synthesize the amylase enzyme utilizing synthetic soluble starch and sugar cane bagasse as substrates. Aspergillus niger, Aspergillus flavus, and Fusarium spp., previously identified to have amylolytic activity, were used for the research. The submerged fermentation process was used to produce amylase, with basal medium and sugar cane bagasse as substrates. The incubation time, substrate and inoculum concentrations, pH, and temperature were all optimized. The dinitrosalicylic acid reagent (DNS) technique was used to determine the activity of the amylase generated. Initial production of amylase using soluble starch (20 g (w/v)) as substrate at room temperature and pH 7.0 shows that all the isolates produce amylase better when their concentration (3%) is high but have different incubation periods of 96 hr for Aspergillus niger (8.65±0.21 U/mL/minutes) and Fusarium spp (7.15±0.07 U/mL/minutes), while Aspergillus flavus (7.30±0.14 U/mL/minutes) requires an extended incubation period of 144 hr to produce the product. Further production using sugar cane bagasse and optimization of production parameters of the isolates reveals that Aspergillus niger (4.35±0.07 U/mL/minutes) has an optimum incubation period of 120 hours, an inoculum concentration and substrate concentration of 2% each, and a pH of 6, Aspergillus flavus (6.40±0.28 U/mL/minutes) has an optimum incubation period of 144 hours, with inoculum and substrate concentration of 3% each at neutral pH, Fusarium spp (6.80±0.28 U/mL/minutes) has an optimum incubation period of 168hr., inoculum concentration of 3%, substrate concentration of 2% and a neutral pH-7 and all the isolates have their best yield at temperature of 30oC. In conclusion, using cheap and readily available sugar cane bagasse can be a more cost-effective option for expensive synthetic starch substrates used in amylase synthesis. The study suggests further investigation into the isolates' identities and the extracted enzyme's industrial application.

A study on synthesis of amylase, cellulase L-asparaginase enzymes and potassium solubulizing efficiency of rhizobacteria from the Piper betle fields of Guntur District, Andhra Pradesh, India

A study on synthesis of amylase, cellulase L-asparaginase enzymes and potassium solubulizing efficiency of rhizobacteria from the Piper betle fields of Guntur District, Andhra Pradesh, India

Light and oxygen facilitating the directly treatment food wastewater and poly-β-hydroxybutyrate, 5-aminolevulinic acid, pigment productions by Rubrivivax gelatinosus.

Rubrivivax gelatinosus has the advantage of using wastewater to realize biomass recovery. However, they still cannot be applied large scale because they cannot directly treat the wastewater containing macromolecular organics. Thus, this article investigated the effects of light-oxygen conditions on R. gelatinosus by directly recycling wastewater containing macromolecular organics to produce biomass, poly-β-hydroxybutyrate (PHB), 5-aminolevulinic acid (5-ALA), and pigment. Results showed that R. gelatinosus directly treated the macromolecule organic (soybean protein and starch) wastewaters and achieved biomass recovery under light-anaerobic and light-micro-oxygen in six conditions. Chemical oxygen demand, protein, and starch removals for two wastewaters all reached above 70%. Renewable bio-resources such as biomass, PHB, 5-ALA, and pigment production were 10 times the initial content. Theoretical analysis indicated that light activated the synthesis of protease and amylase. However, oxygen concentration decided the number of enzymes. When oxygen was at micro-oxygen or anaerobic, the aforementioned expression and synthesis were conducted. In summary, this study expanded the viewpoint ignored by traditional theory. It was realized that R. gelatinosus directly treated wastewater and accumulated nutrients (biomass, PHB, pigment, and 5-ALA) for recycling, which reduced the secondary pollution of excess sludge into the environment.

Farinaceous and starchy foods in the diet of the indigenous people of the high-latitude and Arctic regions of Russia: tradition and modernity

The object of the study is the indigenous population of the North and the Arctic of the Russian Federation. The subject of the study is changes in nutrition during transition from the traditional to modernized lifestyle. The purpose of the study is to consider the risk to the health of the northerners in the quantitative and qualitative changes in the consumption of flour and starch-containing products. The consumption of bread products and starch by the population of the northern regions of the Russian Federation in the 19th – 20th centuries and in the 2010s – 2020s was estimated. Estimates of the frequencies of the amylase genes AMY1, AMY2B, AMY2A in the populations that differ in geographic localization and type of management are systemized. In the 20th century, per-capita consumption of bread in Russia decreased from 700 to 260 g/day, among the indeginous northerners — from 300 to 250 g/day. The inclusion of pasta products and potato in the diet of the northerners changed the volume and structure of the consumed carbohydrate foods. The intake of high glycemic load starch increased from 100 to 250 g/day. Replacing the dried crusts with pasta and potato meals eased mastication of food, thus inducing a reflectory rise in serum glucose at ingestion and increasing the risk of developing chronic carbohydrate metabolism disorders. High-latitude populations are distinct in the prevalence of the genotypes that determine low activity of the polysaccharidase enzymes necessary for starch digestion. The ample production of amylase and less prominent blood glucose upswing in response to the consumption of starchy foods are determined by the number of successive copies of the amylase genes on the AMY locus. For the AMY1 gene, a negative correlation was found between the gene copy number and the geographical latitude of the population area (rSp = -0.19), for the AMY2A gene rSp = -0.33 (p < 0.001 in both cases). In addition, in the Arctic groups there is a high frequency of deletions of the AMY2A gene, which also lead to insufficient synthesis of pancreatic amylase. In the European populations, AMY2A deletions occur in 10–11 %; among the Khanty, Mansi, and Selkups — in 28 %; among the indigenous population of Chukotka — in 52 %. Changes in the “Arctic diet” in the 20th century led to a sharp increase in the starch consumption. With the traditional way of life, the northerners did not form adaptations to the nutrient intake of large volumes of starchy foods. In modern conditions, accumulation of nutritional, physiological and genetic factors increases the risk of metabolic disorders related to the consumption of high-starch food.

Multiple cystic sphere formation from PK-8 cells in three-dimensional culture

Three-dimensional (3D) culture of cancer cells mimics the in vivo environment. Recently, we reported that pancreatic ductal adenocarcinoma (PDAC) cell lines with epithelial and mesenchymal features formed differently shaped spheres in 3D culture. However, only PK-8 cells, the epithelial PDAC cell line with the highest E-cadherin expression among the eight PDAC cell lines, formed multiple cystic spheres in 3D culture. Optical coherence tomography revealed interconnected cysts inside the spheres. A weak inter-cellular adhesion, individual cell degeneration, necrosis, and secretory granules in the cytoplasm were observed in the PK-8 spheres using electron microscopy. The expression of MUC1, MUC5AC, and amylase was increased in PK-8 cells in the 3D culture compared with that in 2D culture. These findings suggest that highly E-cadherin-expressing epithelial PK-8 cells form multiple cystic spheres, which may be promoted by enhanced mucin and amylase synthesis in 3D culture.

Biosynthesis and characterizations of extracellular enzymes of microbial isolates from dairy industrial effluent

Diary effluents are environmental pollutant which could serve as source of substrate for microbial enzymes' synthesis. This research investigated the isolation and screening of bacteria and fungi with potential extracellular proteolytic and amylolytic properties from dairy effluents, and their characteristics for maximum enzyme production. Effluent from a dairy industry located in Ibadan, Nigeria, was collected for isolation and screening of microbial species for enzymatic activities. The Biological Oxygen Demand (BOD), Total Soluble Solid (TTS), and pH of the effluent were determined using standard procedures. The potential isolates were characterized morphologically and by molecular techniques. Proteolytic and amylolytic activities of isolates were investigated on skimmed milk starch agar respectively and optimized with varying pH (6.0 - 9.0), temperature (27 - 60 °C) and incubation period (12 - 72 h). Data were analyzed by Duncan Multiple Range Test. Biological Oxygen Demand, Total Soluble Solid and pH of the effluent were 14.67 mg/L, 13.33% and 7.58 respectively. Eleven isolates were positive for either protease or amylase synthesis, but only two showed both proteolytic and amylolytic activities and were molecularly characterized as Bacillus stratosphericus CM2HG6 and Aspergillus welwitschiae. Optimization showed that enzyme production by B. stratosphericus CM2HG6 was highest at 35 °C, pH 7.0 for 48 h, and that of A. welwitschiae achieved optimum production at 37 °C, pH 8.0 for 72 h. Under optimized conditions, Bacillus stratosphericus CM2HG6 was the highest amylase and protease producer (24.4 and 30.74 U/ml respectively), while A. welwitschiae was the lowest producer (12.58 and 18.8 U/ml respectively). The study successfully showed that these industrially-important enzymes can be produced by microbial strains isolated from dairy effluent, and production can be further optimized

The effect of probiotics on digestive enzyme activity during larvae and juvenile stage of Yellow Fin Tuna (Thunnus albacares)

Yellow fin tuna fry production technology is not successful yet. This was assumed related to the biological and physiological characteristics of the fish. The physiological approach to regulating digestion through probiotics is needs to be studied. The aim of this study was to examine the effect of probiotics bacterial strains of the enzymatic activity in digestion system for the larvae to juvenile stages of yellow fin tuna, Thunnus albacares. The experiment was initiated by culturing 3 probiotic isolates (Bacillus subtilis strain TA-1, Bacillus amyloliquefaciens strain TN-2, Bacillus subtilis strain TO-4) and apply to the larval rearing of yellow fin tuna. In the experiment the treatments applied were supplemented probiotics and without probiotic, each treatment was held with 2 replications. Fish samples were collected every day and analyze the gene expression profile associated with digestion enzyme synthesis by total RNA isolation and cDNA synthesis followed analysis of its by using the RT –qPCR method. The results showed that the growth response of T. albacares tuna larvae reared with probiotics tended to be faster (P<0.05) than the control (without probiotics). The supplementation of probiotic could improved the expressed digestibility of the target genes associated with enzymatic synthesized. Amylase enzymatic synthesized increase 200-1200 times in larvae 10 –17 Day After Hatch (DAH), while in the lipase enzymatic activity started with 13 DAH ( 25 time) and 16 – 21 DAH by 150-300 times. In trypsin the highest enzymatic activity was at 16 DAH, which was 200 times. While in control, the expression of enzymatic synthesis of amylase, lipase, and trypsin was relatively low.

Co-optimization of B. licheniformis 208 biomass and alpha amylase synthesis using response surface methodology

Co-optimization of B. licheniformis 208 biomass and alpha amylase synthesis using response surface methodology

Physiological Studies of Some Insecticides on Spodoptera littoralis Larvae

The present study aims to focus on the effect of the sublethal rates of Lambda-cyhalothrin +Thiamethoxam and Lambda-cyhalothrin, which belonging to groups 3 and 4 on the 4th larval instar of Spodoptera littoralisfield strain under laboratory conditions, and their effect on some physiological aspects, post-treatment with rates 50 and 100 ml/200L for 48 hrs. The results cleared that highly physiological disturbance was recognized as marked stimulating in total protein, total lipids, trehalase, AchE, and acid phosphatase synthesis however, inhibit amylase, invertase and alkaline phosphatase for lambda-cyhalothrin + Thiamethoxam. The tested sublethal rate of lambda-cyhalothrin, also be highly affected by stimulating total protein, invertase, trehalase, AchE, acid and alkaline phosphatases. On the other hand, inhibits total lipids and amylase synthesis in insect cells.

Secrets, puzzles and mysteries of macroamylasemia

Physiological features of amylase synthesis and excretion are considered in the article, presence of other sources of amylase synthesis different from pancreas and salivary glands is emphasized. Definitions of hyperenzymemia and macroamylasemia (MAE) are given. MAE is a state characterized by presence of circulating complexes of normal serum amylase with protein or carbohydrates in blood. There are 3 types of MAE: first — classical (constant hyperamylasemia, decreased amylase level in urine, high blood concentration of macroamylase complexes); second — hyperamylasemia with slightly decreased amylase activity in urine, macroamylase/normal amylase ratio is less than in the first type; third — normal blood and urine amylase activity, low macroamylase/normal amylase ratio. Pathogenesis is explained by connection of blood amylase and acute phase protein in different inflammatory, infectious diseases, malabsorption. MAE clinical manifestations could be absent, sometimes abdominal pain is possible. Hyperamylasemia and reduced urine amylase activity are typical. MAE diagnostics means determination of macroamylase complexes in blood (chromatography, calculation of the clearance ratio of amylase and creatinine). The article presents clinical cases describing extra-pancreatic MAE in women with malignant ovarian lesions. The question of expediency of thorough diagnostic examination in asymptomatic MAE is raised, which may turn out to be a symptom of cancer. The lack of specific treatment for MAE is emphasized.

Entrapment of porous cross-linked enzyme aggregates of maltogenic amylase from Bacillus lehensis G1 into calcium alginate for maltooligosaccharides synthesis

Maltooligosaccharides (MOSs) are emerging oligosaccharides in food-based applications and can be synthesized through the enzymatic synthesis of maltogenic amylase from Bacillus lehensis G1 (Mag1). However, the lack of enzyme stability makes this approach unrealistic for industrial applications. The formation of cross-linked enzyme aggregates (CLEAs) is a promising tool for improving enzyme stability, and the substrate accessibility problem of CLEA formation was overcome by the addition of porous agents to generate porous CLEAs (p-CLEAs). However, p-CLEAs exhibited high enzyme leaching and low solvent tolerance. To address these problems, p-CLEAs of Mag1 (Mag1-p-CLEAs) were entrapped in calcium alginate beads (CA). Mag1-p-CLEAs-CA prepared with 2.5% (w/v) sodium alginate and 0.6% (w/v) calcium chloride yielded 53.16% (17.0 U/mg) activity and showed a lower deactivation rate and longer half-life than those of entrapped free Mag1 (Mag1-CA) and entrapped non-porous Mag1-CLEAs (Mag1-CLEAs-CA). Moreover, Mag1-p-CLEAs-CA exhibited low enzyme leaching and high tolerance in various solvents compared to Mag1-p-CLEAs. A kinetic study revealed that Mag1-p-CLEAs-CA exhibited relatively high affinity towards beta-cyclodextrin (β-CD) (Km = 0.62 mM). MOSs (300 mg/g) were synthesized by Mag1-p-CLEAs-CA at 50 °C. Finally, the reusability of Mag1-p-CLEAs-CA makes them as a potential biocatalyst for the continuous synthesis of MOSs.

Isolation and purification of starch hydrolysing amylase from Streptomyces sp. Al-Dhabi-46 obtained from the Jazan region of Saudi Arabia with industrial applications

Amylase is an important industrially useful enzyme with various applications, widely explored for their applications in food and biofuel industries. In this study, seventeen actinomycetes strains were isolated from the soil sample and screened for amylase biosynthesis. Among the 17 isolates, nine actinomycetes showed activity on starch agar plates. The strain Al-Dhabi-46 showed more hydrolytic zone (19 mm) on starch agar plates. It was identified by biochemical characters and 18S rDNA sequencing and characterized as Streptomyces sp. Al-Dhabi-46. Amylase production was found to be maximum after 5 days of incubation (108 ± 4.1 U/ml) and at pH 8.0 (118.1 ± 2.3 U/ml) and at 40 °C (124 ± 12.1 U/ml). Amylase synthesis was high in the medium containing starch (208 ± 11.4 U/ml). Amylase production was 241 ± 18.1 U/ml in the culture medium supplemented with beef extract and 0.1% Mn2+ ions significantly enhanced amylase production. The molecular weight of purified amylase from Streptomyces sp. Al-Dhabi-46 was 44 kDa. Amylase was highly active up to pH 8.0 and at 40 °C. Amylase activity was enhanced by the addition of Mn2+ at 10 mM concentrations. Purified amylase from Streptomyces sp. Al-Dhabi-46 digested cassava starch at 25% and 55%, 78% and 95.4% level after 15 min, 30 min, 45 min and 60 min of enzymatic reaction. Enhanced production of amylase enzyme from the Streptomyces sp. Al-Dhabi-46 attract for the production of industrially important products.

Isolation and characterization of a cold-active, alkaline, detergent stable α-amylase from a novel bacterium Bacillus subtilis N8

ABSTRACTA cold-active alkaline amylase producer Bacillus subtilis N8 was isolated from soil samples. Amylase synthesis optimally occurred at 15°C and pH 10.0 on agar plates containing starch. The molecular weight of the enzyme was found to be 205 kDa by performing SDS-PAGE. While the enzyme exhibited the highest activity at 25°C and pH 8.0, it was highly stable in alkaline media (pH 8.0–12.0) and retained 96% of its original activity at low temperatures (10–40°C) for 24 hr. While the amylase activity increased in the presence of β-mercaptoethanol (103%); Ba2+, Ca2+, Na+, Zn2+, Mn2+, H2O2, and Triton X-100 slightly inhibited the activity. The enzyme showed resistance to some denaturants: such as SDS, EDTA, and urea (52, 65, and 42%, respectively). N8 α-amylase displayed the maximum remaining activity of 56% with 3% NaCl. The major final products of starch were glucose, maltose, and maltose-derived oligosaccharides. This novel cold-active α-amylase has the potential to be used in the industries of detergent and food, bioremediation process and production of prebiotics.

Predicting enzymatic starch hydrolysis mechanism during paddy malting by vibrational spectroscopy and multivariate calibration analysis

Vibrational spectroscopic techniques were employed to predict the mechanism of starch hydrolysis based on structural changes during germination of paddy. The proposed mechanism for starch hydrolysis dealt with the synthesis of amylase at the onset of germination, depicting an increased intensity of spectral bands at amide I, II and III regions. The process commenced with the enzyme actions on skeletal mode of pyranose ring structure of glucose units followed by cleavage of the glycosidic linkage by the process of multiple and multi-chain attack resulting in decrease of the bands (400–900 cm−1). The increased intensity of the bands (1200–1500 cm−1) indicated the process of starch hydrolysis and formation of d-glucose. Multivariate calibration analysis (PCA and PLS) was employed to correlate Raman spectral data with biochemical changes during germination and to develop a calibration model. The model showed a high prediction ability with low root mean square error of prediction (RMSEP) (0.043–0.568).

Induction and catabolite repression of alpha-amylase synthesis in Bacilluslicheniformis SKB4

Microbial amylases have an exciting potentiality and are being used extensively in different industries. In this study, regulation of amylase biosynthesis was examined in Bacillus licheniformis SKB4 (wild type) and its mutant strain (8b). The mutant strain was developed by using UV exposure. Expression of the a-amylase gene of Bacillus licheniformis was activated by inducer and subject to catabolite repression. Addition of exogenous glucose or sucrose repressed bio-synthesis of a-amylase which was concentration (0.05-1.0% w/v) dependent. However, mutant strain could enable to overrule the glucose mediated repressive effect. Supplementation of second messenger like cyclic adenosine 3',5'-monophosphate (cAMP, 5 mM) along with glucose could a little bit improve amylase synthesis in wild strain. Antibiotics like rifampicin and tetracycline (ribonucleic acid and protein synthesis inhibitor; 100mg/ml) had stopped the release of enzyme in both wild and mutant strain. Amylase production was also inhibited in presence of respiratory inhibitor 2,4-dinitrophenol (uncoupler) at (5mM) concentration. Thus, the pattern of regulation of a-amylase production in the present strain was in multiple forms; it showed the classical glucose effect without stimulation of second messenger system.

RCAD/BiP pathway is necessary for the proper synthesis of digestive enzymes and secretory function of the exocrine pancreas.

Alcoholism causes an imbalance of endoplasmic reticulum (ER) homeostasis in pancreatic acini. In those cells, the ER is involved in the synthesis and folding of pancreatic enzymes. Ubiquitin-fold modifier 1 (Ufm1) is part of a novel ubiquitin-like modification system involved in maintaining ER homeostasis. Among the components of the Ufm1 system, Regulator of C53 and DDRGK1 (RCAD) has recently been identified as a Ufm1-specific E3 ligase that promotes ufmylation of DDRGK1, an RCAD-interacting protein. We determined the importance of RCAD in the proper synthesis and secretion of pancreatic enzymes using mice with genetically deleted RCAD. The pancreas of RCAD-deficient mice was of normal size and histology. Using quantitative PCR and Western blotting, we found that amylase was upregulated in pancreas organs from RCAD-knockout (KO) mice. Constitutive amylase secretion was much higher in isolated pancreatic acini from RCAD KO mice, whereas CCK-stimulated amylase secretion was disturbed. RCAD deficiency caused a downregulation in expression of ER chaperone BiP, which affected ER homeostasis and activated both apoptosis and trypsin. We also found that both RCAD and DDRGK1 transcript levels were upregulated in pancreatic acini from alcohol-preferring rats. Elevated expression of RCAD and DDRGK1 was associated with increased ER stress and UPR activation. Because of the lack of BiP expression, caspase 3 and trypsin activation we enhanced in RCAD-deficient pancreatic acini upon treatment with ethanol and CCK. In conclusion, the RCAD/BiP pathway is required for proper synthesis and secretion of pancreatic enzymes. In alcoholism, increased levels of components of the Ufm1 system could prevent the deleterious effects of alcohol in the pancreas by regulating BiP levels.NEW & NOTEWORTHY RCAD/BiP pathway is required for the proper synthesis and secretion of amylase from pancreatic acini, as well as for the maintenance of the ER homeostasis. In alcoholism, the exocrine pancreas could increase the levels of components of the Ufm1 system to protect itself from alcohol's deleterious effects by regulating the expression of ER chaperone BiP.

Simultaneous production of amylases and proteases by Bacillus subtilis in brewery wastes.

The simultaneous production of amylase (AA) and protease (PA) activity by Bacillus subtilis UO-01 in brewery wastes was studied by combining the response surface methodology with the kinetic study of the process. The optimum conditions (T = 36.0 °C and pH = 6.8) for high biomass production (0.92 g/L) were similar to the conditions (T = 36.8 °C and pH = 6.6) for high AA synthesis (9.26 EU/mL). However, the maximum PA level (9.77 EU/mL) was obtained at pH 7.1 and 37.8 °C. Under these conditions, a considerably high reduction (between 69.9 and 77.8%) of the initial chemical oxygen demand of the waste was achieved. In verification experiments under the optimized conditions for production of each enzyme, the AA and PA obtained after 15 h of incubation were, respectively, 9.35 and 9.87 EU/mL. By using the Luedeking and Piret model, both enzymes were classified as growth-associated metabolites. Protease production delay seemed to be related to the consumption of non-protein and protein nitrogen. These results indicate that the brewery waste could be successfully used for a high scale production of amylases and proteases at a low cost.

Insulin and alpha amylase levels in alloxan-induced diabetic rats and the effect of Rothmannia hispida (K. Schum) Fagerl leaf extract

Objectives: Rothmannia hispida (R. hispida) is used in West African traditional medicine for the treatment of various ailments such as fever, dysentery, skin infections, abdominal pain and diabetes mellitus. To elucidate the pharmacological basis of the antidiabetic efficacy of this plant drug, the effect of R. hispida on insulin and alpha amylase levels were examined in alloxan-induced diabetic rats. Method: Diabetic rats were treated with leaf extract of R. hipida at dose levels of 250mg/kg and 500mg/kg respectively. The concentration of insulin in serum samples was estimated using Enzyme-Linked Immunoabsorbent Assay (ELISA) method using insulin kit (Syntron Bioresearch, USA), alpha amylase levels were estimated using routine biochemical procedures. Results: Treatment of alloxan diabetic rats with leaf extract of R. hipida significantly (p&lt;0.001) reduced hyperglycaemia, significantly (p&lt;0.001) attenuated alloxan-induced hypoinsulinaemia and significantly (p&lt;0.01) increased alpha amylase levels compared with diabetic untreated rats. Conclusion: It is concluded that increased insulin secretion and/or increased alpha amylase synthesis sequel to enhanced liver glucose entry by Rothmannia hispida is proposed to be the mechanism by which this herbal plant exhibits its antidiabetic effect.