Maximum Amylase Research Articles

Production of Amylase by Solid State Fermentation Using Agricultural Waste

This study presents a comprehensive investigation into the production of amylase, a crucial enzyme with wide-ranging industrial applications, using locally sourced substrates from Kachchh, Gujarat. The research employed the Bacillus licheniformis strain and substrates such as coconut, rice husk, wheat bran, paddy straw, and maize straw. The study found paddy straw to be the most promising substrate for amylase production. The research also systematically optimized various process parameters for amylase production in Solid-State Fermentation (SSF) using the One Variable at a Time (OVAT) method. These parameters included incubation period, temperature, inoculum level, additional carbon sources, starch concentrations, additional nitrogen sources, initial pH, different mineral salt ions, initial moisture level, and surfactants. The results showed that the optimal conditions for maximum amylase yield were an incubation period of 48 hours, an incubation temperature of 35°C, an inoculum level of 10%, starch as the additional carbon source, a starch concentration of 2.5%, yeast extract as the additional nitrogen source, an initial pH of 7, NaCl as the mineral salt, an initial moisture level of 75%, and Tween 80 as the surfactant. This research provides a reliable and sustainable approach to enzyme production, offering valuable insights for the optimization of the solid-state fermentation process for maximum amylase production.

Examining surgeon stress in robotic and laparoscopic surgery.

Robotic surgery may decrease surgeon stress compared to laparoscopic. To evaluate intraoperative surgeon stress, we measured salivary alpha-amylase and cortisol. We hypothesized robotic elicited lower increases in surgeon salivary amylase and cortisol than laparoscopic. Surgical faculty (n = 7) performing laparoscopic and robotic operations participated. Demographics: age, years in practice, time using laparoscopic vs robotic, comfort level and enthusiasm for each. Operative data included operative time, WRVU (surgical "effort"), resident year. Saliva was collected using passive drool collection system at beginning, middle and end of each case; amylase and cortisol measured using ELISA. Standard values were created using 7-minute exercise (HIIT), collecting saliva pre- and post-workout. Linear regression and Student's t test used for statistical analysis; p values < 0.05 were significant. Ninety-four cases (56 robotic, 38 laparoscopic) were collected (April-October 2022). Standardized change in amylase was 8.4 ± 4.5 (p < 0.001). Among operations, raw maximum amylase change in laparoscopic and robotic was 23.4 ± 11.5 and 22.2 ± 13.4; raw maximum cortisol change was 44.21 ± 46.57 and 53.21 ± 50.36, respectively. Values normalized to individual surgeon HIIT response, WRVU, and operative time, showing 40% decrease in amylase in robotic: 0.095 ± 0.12, vs laparoscopic: 0.164 ± 0.16 (p < 0.02). Normalized change in cortisol was: laparoscopic 0.30 ± 0.44, robotic 0.22 ± 0.4 (p = NS). On linear regression (p < 0.001), surgeons comfortable with complex laparoscopic cases had lower change in normalized amylase (p < 0.01); comfort with complex robotic was not significant. Robotic may be less physiologically stressful, eliciting less increase in salivary amylase than laparoscopic. Comfort with complex laparoscopic decreased stress in robotic, suggesting laparoscopic experience is valuable prior to robotic.

BACTERIAL AMYLASE PRODUCTION BY THE L-LACTIC ACID-PRODUCING STRAIN USING CASSAVA STARCH SUPPLEMENTED WITH RICE BRAN

Amylase is a crucial starch-degrading enzyme, which is commercially available, prvides benefit to starchy industry to overcome viscosity problem that caused by using high starch concentration, and to supply sugar residues. This study aimed to investigate the production of amylase by the potential optical pure L-lactic acid-producing Lactococcus sp. SUT 513 for further development of the optical pure acid fermentation process from the cheap cassava starch. Rice bran, the main byproduct in the process of rice milling, was found to provide the maximum amylase production when supplemented with 4.0 g/L (w/v) to the medium containing cassava starch (20.0 g/L, w/v), compared to the cassava medium added either the expensive nitrogen source, tryptone (5.0 g/L) and yeast extract (3.0 g/L, w/v); or the cheap alternatives, soy ptotein (8.0 g/L) or urea (2.0 g/L) combined with yeast extract (3.0 g/L). The amylase activity obtained from the developed medium was approximately 3 times higher than the original medium. This bacterium gave the highest crude amylase activity of 4.848±0.237 U/mL in the suitable medium performed in a7.5-L bioreactor with 3.0-L working volume for the optimal cultivation time of only 18 h under a control pH of 7.0, temperature at 35°C, agitation at 150 rpm, and without aeration. The results reveal the high potential production of amylase by Lactococcus sp. SUT 513 using the cheap cassava starch medium supplemented with rice bran, providing an opportunity to develop both amylase and optically pure L-lactic acid production processes.

A Novel Halo-Acid-Alkali-Tolerant and Surfactant Stable Amylase Secreted from Halophile Bacillus siamensis F2 and Its Application in Waste Valorization by Bioethanol Production and Food Industry.

The extracellular amylase production level by the moderate halophile Bacillus siamensis F2 was optimized, and the enzyme was biochemically characterized. The culture parameters for NaCl, carbon, nitrogen, pH, and temperature were optimized for high titers of amylase production. Growing B. siamensis F2 cultures in Great Salt Lake-2 medium with additions of (in g/L) NaCl (100), starch (30), yeast extract (2), KNO3 (2), and MgSO4 (1) at pH 8, 30 °C resulted in the maximum amylase production (4.2 U/ml). The amylase was active across a wide range of salinities (0 to 30% NaCl), pH (5.0-10.0), and temperatures (20-70 °C) and showed good stability with surfactants (sodium dodecyl sulfate (SDS) and Triton X-100); hence, it was identified as halo-acid-alkali-tolerant and surfactant stable. Temperature, pH, and salinity were optimal for amylase activity at 50 °C, pH 7, and 5% NaCl, respectively. It also generates amylase by utilizing agricultural wastes like sugarcane bagasse, sweet potato peel, and rice husk. Based on the performance of B. siamensis F2 using agricultural wastes and synthesizing amylase, the current study attempted to produce bioethanol by coculturing with baker's yeast using sugarcane bagasse and sweet potato peel as a substrate, which yielded 47 and 57 g/L of bioethanol, respectively. Besides bioethanol production, amylase secreted by F2 was also employed for juice clarification for better yield and clarity and for softening dough to produce better-quality buns. This novel amylase may have many potential applications in waste valorization, biorefinery sectors, and food industries.

LONG-TERM LIGHT DEPRIVATION AFFECTS DIGESTIVE FUNCTION IN RATS DURING ONTOGENY

The effects of light deprivation on age-related changes in body weight, food and water intake, as well as the activity of digestive enzymes in pancreatic tissues of male rats was studied. Animals were divided into three groups: the first was in standard light conditions (12 h light/12 h dark, control, LD), the second was kept under conditions of long-term light deprivation from the moment of birth (LD/DD), and the third – from the prenatal period (DD/DD). Prolonged keeping of rats under conditions of light deprivation led to disruption of the age-associated dynamics of the studied parameters, at the same time, the detected changes in response to a specific light condition differed depending on the stage of ontogenesis at which its exposure began. Thus, body mass (age 3 and 18 months) and food and water intake (12 months), amylase and lipase activities (18 months) were lower, and protease activities after the end of the stable growth stage (12, 18 and 24 months) were higher in LD/DD-rats than in LD-rats. Significant changes in the studied parameters were observed under DD/DD conditions in 12‑month-old and 18-month-old rats – body mass was the largest, and food and water intake were the lowest compared to LD and LD/DD. In addition, the maximum amylase activity (1 and 6 months) among the studied groups and the higher values of total proteolytic activity during the period of stable growth (6 and 12 months) were found in DD/DD-rats compared to controls. Thus, our results indicate that light deprivation disrupts the ontogenetic development program of the digestive system of mammals.

Efficacy of the Fruit and Vegetable Peels as Substrates for the Growth and Production of α-Amylases in Marine Actinobacteria.

Enzymes from haloalkaliphilic microorganisms have recently focused attention on their potential and suitability in various applications. In this study, the growth and production of extracellular amylases in the marine actinomycetes, using kitchen waste as the raw starch source, have been investigated. Actinobacteria were isolated from the seawater of the Kachhighadi Coast near Dwarika, Gujarat. Seven Actinobacterial isolates of pre-monsoon, monsoon, and post-monsoon seasons belonging to different strains of Nocardiopsis genera were screened and selected for amylase production. The amylase production was initially assessed on the solid media supplemented with the extracts of different fruits and vegetable peels as a substrate by agar plate assay. The strains Kh-2(13), Kh-2(1), and Kh-3(12) produced maximum amylase with potato peel as a substrate, while no significant differences were found with the media containing other peels. Nevertheless, all strains produced amylases at a significant level with other raw substrates as well. For the optimization of the growth and enzyme production, the selected two isolates Kh-2(13) and Kh-3(12) of the monsoon and winter seasons were cultivated in a liquid medium under the submerged fermentation conditions, with potato peel as a substrate. In both organisms, the optimum amylase production was observed in the stationary phase of growth. For amylase production, the effect of different physical and chemical parameters was evaluated. The optimum growth and amylase production was achieved in 2% inoculum size, at pH 8.0, 28℃, and 5% salt concentration. On the basis of the amylase production index (API) (a ratio of the amylase units and cell growth), both isolates produced significant amylase with the only extract of potato peels, without any other supplements. The trends further indicated that while additional complex sources, such as yeast extract and peptone can enhance the cell growth of the actinobacteria, the amylase production remained unaltered. The study projects the significance of waste raw materials for the production of enzymes in extremophilic microorganisms.

Valorization of banana peel into α-amylase using one factor at a time (OFAT) assisted artificial neural network (ANN) and its partial purification, characterization, and kinetics study

In this present study α-amylase producing bacteria were isolated from the soil near North Bengal University canteen on an amylase agar medium. Primary screening of the isolates was done by iodine test, based on the clear zone around the bacterial colonies in the media plates. Seven bacterial isolates were selected and quantitatively screened for amylase production. Among them, strain sps2 showing the highest amylase activity (215 ± 2 IU/ml) was selected for further studies. Isolated strain sps2 was identified both morphologically and biochemically and further confirmed by 16s rRNA sequencing as Streptomyces pratensis sps2 (Gene bank accession No. OP 236721). Optimization of α-amylase produced by the isolate was conducted under submerged fermentation of low-cost carbon source banana peel, using a one factor at a time (OFAT) approach. Maximum amylase activity of 821.33 ± 0.57 IU/ml was recorded after 2 days of fermentation. The artificial neural network (ANN) model was employed using the OFAT data to generate a predictive model for amylase production and the R-value of 0.94496 and the RMSE of 64.82 suggesting that the neural network has the precise capability to generate an optimization environment. Ammonium sulfate precipitation and dialysis techniques were used to partially purify the α-amylase and then characterized. The enzyme was found to have a molecular mass of 28 kDa (SDS-PAGE and zymogram) with a Km and Vmax of 2 mg/ml and 1000 μmol/min, respectively. The enzyme was found to be thermostable in the temperature range of 4–90 °C and more stable at alkaline pH with optimum pH of 7.

Production of Amylase from the fungus Aspergillus niger FA6 Isolated from Iraqi Soil.

Filamentous fungi have been widely used to produce hydrolytic&#x0D; enzymes for many applications. Fungal enzymes amylase were&#x0D; produced using eight Aspergillus niger isolates from eight soil sample in&#x0D; Iraq, were screened for amylase production. A. niger FA6 (isolated from&#x0D; AL-Nasiriya soil) gave maximum amylase yield, The effect of varying&#x0D; &#x0D; pH, temperature, incubation period, carbon sources and_ nitrogen&#x0D; sources of the medium for the productivity of amylase from&#x0D; A.nigerFA6 was _ investigated. The maximum activity of amylase was&#x0D; recorded after 7 days of submerged fermentation at pH 5 and temperature&#x0D; 30°C. from different nitrogen and carbon sources used peptone and&#x0D; sucrose respectively gave maximum amylase yield.

Optimization of α-Amylase Production by Enterobacter cloacae Strain D1 Isolated from Cassava Effluent-impacted Soil Using Response Surface Methodology

Background: Production of amylase by Enterobacter cloacae D1 was optimized in this study using central composite design (CCD) of response surface methodology (RSM).&#x0D; Methodology: Effects of five numeric factors (pH, temperature, inoculum concentration, peptone and yeast extract) on the production of amylase were examined. Amylase production was first screened using plate technique and amylase assay thereafter carried out using the dinitrosalicylic acid (DNSA) method. The CCD-RSM experimental set-up involved 30 runs with 5 levels of independent variables.&#x0D; Results: The amylase-producing bacterium Enterobacter cloacae strain D1 was identified based on the phylogenetic tree analysis of its sequence. The sequence has been submitted to GenBank under the accession number: MZ477010. The isolate had 98% similarity to the GenBank match Enterobacter cloacae strain ATCC 13182. Optimum conditions that yielded maximum amylase (34.43 U/mL) were pH 5; temperature 40 ℃; inoculum concentration 3%; peptone 1.2% and yeast extract 0.5%.&#x0D; Conclusion: This study has demonstrated efficient amylase production from Enterobacter cloacae strain D1 isolated from cassava effluent-impacted soil from Rumuosi, Port Harcourt, Rivers State. In addition, optimization of the critical factors of amylase production resulted in 3.4 fold increase in amylase activity. The enhancement of amylase production by the RSM techniques shows that amylase from this strain can be scaled-up for industrial application.

Production of lactic acid from pasta wastes using a biorefinery approach

A total of 398 kt of pasta waste (PW), generated during the production process of pasta, were produced in 2021. Due to its chemical composition and practically zero cost, PW has already been studied as a raw material for the production of lactic acid (LA) through fermentations. The main objective of this article was to improve the economic viability of the process by replacing commercial enzymes, necessary for starch hydrolysis in PW, with raw enzymes also produced from wastes. Enzyme synthesis was achieved through solid-state fermentation (SsF) of wheat bran by Aspergillus awamori or Aspergillus oryzae at various moisture contents. The maximum amylase activity (52 U/g dry solid) was achieved after 2 days of fermentation with A. awamori at 60% of moisture content. After that, the enzymes were used to hydrolyse PW, reaching 76 g/L of total sugars, 65 g/L of glucose and a yield of 0.72 gglu/gds with the enzymes produced by A. awamori. Subsequently, the hydrolysate was fermented into LA using Bacillus coagulans A559, yielding 52 g/L and 49 g/L with and without yeast extract, respectively. Remarkably, compared to the process with commercial enzymes, a higher LA yield was reached when enzymes produced by SsF were added (0.80 gLA/gglu). Furthermore, the productivities between the two processes were similar (around 3.9 g/L/h) which highlights that yeast extract is not necessary when using enzymes produced by SsF.

Production and characterization of α-amylase from indigenously isolated Streptomyces sp.

Streptomyces species have been exploited widely as microbial cell factories, especially for antibiotic production. However, their potential for alpha-amylase production has not been extensively studied. This study reports the isolation, molecular identification, and optimization of the physiological conditions for alpha-amylase production from Streptomyces sp., isolated from soil in Kotli Azad Kashmir. The maximum growth of Streptomyces MI-1 was observed at a neutral pH and a temperature of 35 °C. An amylase activity of 1.15 IU/mL was observed when 4% starch was added to the nutrient medium. During the submerged state fermentation, the maximum amylase activity of 2.136 IU/mL/min was observed after 144 h of incubation. The characterization of bacterial amylase revealed an optimum temperature of 40 °C, and its optimum pH was 7.0. Furthermore, during the study, examining the effect of different metal ions found that Mg2+ and Ca2+ ions had a positive effect, while Cu2+ and Fe2+ ions had an inhibitory effect compared to the control. This preliminary study provides basal line information for the discovery of novel microbes from the unexplored natural resources for amylase production, which will be used for many purposes.

Effect of Incubation Time on Amylase Enzyme Activity in Sticky Rice Tape

Indonesia uses fermentation as a common method of food processing. Tape ketan is one of the foods produced using the fermentation process. The enzyme amylase, which hydrolyzes carbohydrates (starch) into sugar, is involved in the fermentation process of sticky rice tape. Many factors affect the activity of the amylase enzyme. Incubation time is one of them. The length of incubation can also impact the tape's sweetness, color, texture, and scent. The 48- hour incubation period had the maximum amylase activity. Organoleptic testing, meanwhile, revealed a range of color, scent, texture, and taste. We'll talk about how the amylase enzyme's activity and incubation period relate to the color, flavor, texture, and aroma of sticky rice tape in this article

Isolation and characterization of alkaline amylase producers from the marine environment of Arabian sea coast

Alkaline amylase producers were isolated from the marine environment of Arabian Sea coast. A total of 10 isolates were obtained from the soil and water samples collected from the marine environment. Five isolates namely; BCSS, JBWS, RBWS, BCWS and ALSS showed good growth on alkaline M9 media with 10% NaCl having starch as the primary carbon source. Based on their maximal amylase activity, three microbial cultures (BCSS, JBWS, and RBWS) were selected for the study. The crude enzyme from these purified cultures was extracted and partial characterization was carried out. The partial characterization study involved the effect of metal ions and surfactants and determination of optimal pH and temperature and its stability studies. The amylase activity was optimum for all three cultures at 60 0C whereas, the optimum pH was found to be pH 7, pH 8 and pH 9 for RBWS, JBWS and BCSS. The pH and temperature stability studies showed maximum amylase activity at its optimal conditions for 5h (300 mins) in the order BCSS&gt;RBWS&gt;JBWS. The amylase activity was found to increase with sodium (BCSS), manganese (RBWS) and potassium (JBWS) at 1:1 dilution. The cultures were stable with the surfactants (Tween -20, SDS and Triton-X) at 1:10 dilution of 1% surfactant and enzyme. BCSS was the most stable at high temperature and pH among all the isolates with an optimal amylase activity at pH 9 and temperature of 60 0C. The isolate was identified to be Bacillus nanhaiisediminis NH3. These remarkable properties indicated its great potential in industrial applications.

Biotechnologically Important Enzyme Producing Indigenous Bacteria Isolated from Fruit and Vegetable Wastes Samples Collected from different Local Markets

Enzyme from bacterial sources is much stable and obtained cheaply. Amylases and proteases are among the most important enzymes. During this study, indigenous amylase and protease producing bacteria were isolated from common decayed fruits and vegetable wastes viz. Papaya, Brinjal, Cucumber, Potato and Snake Gourd. Bacterial load ranged in between 0.67 × 109 and 9.06 × 109 cfu/g on NA and 1.50 × 109 and 7.00 × 109 cfu/g on PYG agar medium. Maximum mean bacterial load on both NA (6.19 ± 2.60 × 109 cfu/g) and PYG agar (4.54 ± 1.03 × 109 cfu/g) were observed in decayed papaya. A total of 113 bacterial isolates were primarily isolated. Considering better amylase and protease activity 16 isolates were selected for detailed study. The starch hydrolysis ratio (SHR) of the isolates ranged in between 1.25 ± 0.37 and 2.47 ± 0.23 while casein hydrolysis ratio (CHR) ranged in between 2.35 ± 0.12 and 6.44 ± 1.16. The highest SHR was 2.47 ± 0.23 found in Bacillus sp. of snake gourd and the highest CHR was 6.44 ± 1.16 found in Bacillus subtilis also from snake gourd. Out of the 16 isolates 13 were Gram positive and 3 were Gram negative. Gram positive isolates were identified as Bacillus acidocaldarius (4), B. firmus (2), B. lentus (4), B. subtilis (2) and B. alcalophilus (1). Gram negative isolates were identified as Edwardsiella hoshinae, Proteus myxofaciens and P. mirabilis. Six isolates having higher SHR and CHR were authenticated through molecular identification and were identified as Chryseobacterium sp. S29.2, Bacillus sp. X8, Bacillus sp. strain GA1B, Bacillus sp.TdEND26, Bacillus subtilis strain BPA28 and Bacillus subtilis BAB-881. The maximum amylase (61.33 ± 2.14 U/ml) and protease (56.91 ± 0.23 U/ml) production were observed in Bacillus sp. TdEND26 rd. In case of co-production, the highest amount of amylase (54.13 ± 1.23 U/ml) protease (81.80 ± 4.54 U/ml) production was observed in Bacillus sp. TdEND26 at 24 hrs. Plant Tissue Cult. &amp; Biotech. 32(1): 31-41, 2022 (June)

Fermentation optimization and amylase activity of endophytic Bacillus velezensis D1 isolated from corn seeds

To acquire quality amylase adopted in practical applications, endophytic bacteria were identified as Bacillus velezensis strain D1 which was isolated from corn seeds. The fermentation conditions and amylase properties of the strain were investigated. The strain D1 was identified via morphological, physiological and 16S rDNA phylogenetic analysis. The fermentation conditions of secreting amylase were optimized by single-factor and orthogonal experiments. The α-amylase gene was expressed in E. coli and purified by means of immobilized metal ion affinity chromatography (IMAC), upon which the enzyme activity of purified recombinant α-amylase was determined. The results outlined that (1) The strain D1 was identified as Bacillus velezensis. (2) The optimized fermentation conditions for maximum amylase yields included 44°C for 48h at pH 7.5. (3) The enzyme had an optimal reaction temperature of 60°C with the highest activity at 50°C and tolerance to 4-h incubation at 70°C. (4) The enzyme was strong acid resistant and tolerated at pH 5.0-6.0 while the optimal pH was 8.0. (5) Besides, the amylase activity was elevated by the presence of Ca2+ and Cu2+ . (6) The activity of purified recombinant amylase was 20.59U/ml under optimal conditions, nearly seven times that of crude amylase preparations. The amylase produced by Bacillus velezensis D1 is strongly tolerant towards acid and high temperatures. Amylases with thermophilic and acid-resistant characteristics are useful for a wide range of applications in food, brewing, textile, starch, paper and deterrent industries. The enzyme from Bacillus velezensis D1 can be effectively used in different areas of industries.

ISOLATION, CHARACTERIZATION AND EXPLORATION OF THE BIOLOGICAL ACTIVITY OF ENDOPHYTIC BACTERIA FROM MEDICINAL PLANTS

Bacterial endophytes are known to reside inside tissues of plants and can form a variety of different relationships including symbiotic, mutualistic, commensalistic and ammensalism. There has been increasing evidence, that endophytic bacteria can stimulus plant growth significantly by the production of phytohormones analogous to PGPR activity. In the present study, leaf explants of 6 medicinally important plants viz. Abrus precatorius (AP), Aegle marmelos (AM), Aloe barbadensis (AB), Annona squamosal (AS), Azadirachta indica (AI) and Oscimum sanctum (OS) were used for isolation and characterization of bacterial endophytes via morphological, biochemical examinations. Total 21 endophytes were isolated on Luria-Bertani (LB) and Nutrient agar isolation medium. Isolates were further screened for the enzymatic activity (amylase, protease, cellulase and lipase), and antifungal potential against plant fungal pathogen Aspergillus niger. Endophytic bacteria isolates from Aegle marmelos, Aloe barbadensis, and Azadirachta indica showed maximum amylase, protease, cellulase, and lipase activities in addition to antifungal activities. The screened endophytic isolates will be useful as a possible source of novel bioactive metabolites and potential biopesticides and biofertilizers.

Co-culture of Bacillus amyloliquefaciens and recombinant Pichia pastoris for utilizing kitchen waste to produce fengycins

Kitchen waste (KW) is a vast potential source of fermentable substrates. To bio-convert the KW into high-value chemicals, we used KW as substrate for the production of fengycin by an artificial consortium containing Bacillus amyloliquefaciens HM618 producing fengycin and the engineering Pichia pastoris producing amylase, glucosidase, or lipases. The maximal amylase activity of the constructed amylase-producing engineering strain (recombinant P.pastoris GS115-amy98) reached 385.4 U‧mL-1. The engineering strain GS115-α-glu53 producing glucosidase reached an enzyme activity titer of 247.3 U‧mL-1, while the lipase activities of the engineering strains GS115-lip2, GS115-α-lip2, and GS115-lip7 were around 90.0 U‧mL-1, with no significant differences among them. Liquid chromatography-mass spectrometry (LC-MS) analysis showed that the components of fengycin synthesized by B.amyloliquefaciens HM618 were complex, including C14-C18 fengycins A, C13-C14 fengycins B, C16-C18 fengycins B, C16 fengycin B2 and some fengycin homologues with unsaturated fatty acid chains. The levels of fengycin were 15.9mg‧L-1 and 4.6mg‧L-1 under the co-culture with strain HM618 and the recombinant strains producing amylase and lipase, respectively. The maximal titer of fengycin was 21.2mg‧L-1 in the artificial consortia consisting of HM618 and the engineering strains producing glucosidase, amylase and lipase. Taken together, these results show that the co-culture of B.amyloliquefaciens HM618 and engineering strains producing amylase and lipase can promote the conversion of KW into fengycin. The work provides a new strategy for boosting the resource utilization of KW.

Optimization of Amylase Production from &amp;lt;i&amp;gt;Lactobacillus plantarum&amp;lt;/i&amp;gt; CS by Submerged Fermentation Using Agro Wastes Substrates

Amylases are known to be of great importance worldwide. Most of their production using refined carbon and nitrogen substrates are expensive, resulting in the production of expensive goods which are not easily affordable by end users in our country Nigeria. This necessitates the call for the use of cost-effective substrates under optimized condition for maximum amylase yield. Hence, the present study was focused on the optimization of amylase production from <i>Lactobacillus plantarum</i> CS using agro wastes from local substrates. The parameters assayed were carbon and nitrogen substrates from agro wastes, fermentation period (24-168h), pH (3.5-11) and temperature (30°C-70°C). Others included solvents for preparation of fermentation medium and metal ions of Mg²⁺, Mn²⁺, K⁺, Na⁺, Hg²⁺, Fe²⁺, Cu²⁺ and Pb²⁺ as supplements. The amylase produced via shake-flask fermentation was assayed using Dinitro salicylic acid method. The result revealed 3% sweet potato and 1% bambara nut wastes as the best carbon and nitrogen components of the required fermentation medium of pH 6.5. The optimal amylase produced (38.6U/ml) was achieved within 48h at 35°C with 1% inoculum and Mn²⁺/ Mg²⁺ as a co-factor. Relatively, Cu²⁺, Pb²⁺ and Hg²⁺ reduced the amylase yield to < 68.13% as against Mn²⁺ and Mg²⁺ which enhanced the production to 121.66% and 101.09% respectively. Natural rain water used as solvent for the preparation of fermentation medium significantly enhanced amylase production (24.65 U/ml) in relation to other test waters at P>0.05. Comparatively, there was a significant increase in amylase produced under optimized condition than not. Conclusively, the obtained results which revealed high amylase yield with available and cost effective agro-wastes is promising for industrial application in Nigeria.

Polysaccharide hydrolyzing enzyme activity of bacteria, native to Apis florea gut

Introduction and Aim: Apis florea commonly known as “dwarf honey bee” harbors enormous gut bacteria that can digest complex carbohydrates and other food components. In this regard, the present investigation was focused on analyzing the polysaccharide degrading ability of bacteria isolated from the gut of honeybee, for their possible application in nutraceutical and pharmaceutical industries. Materials and Methods: Nine bacterial isolates were screened for carbohydrate degrading enzymes viz., amylase, pectinase, cellulase, tannase and laccase, using respective substrate by plate assay method. Further activities of amylase and pectinase were measured quantitatively by dinitrosalicylic acid (DNS) method. Results: All the nine selected isolates exhibited amylase and pectinase activities. However, only two isolates exhibited lignolytic and cellulolytic activity. None of the isolates showed tannin degradation. Maximum amylase activity (4.95 U/mg) was observed in Bacillus halotolerans af-M9 followed by Klebsiella oxytoca af-G4 (4.62 U/mg). With respect to pectinase activity Klebsiella pneumoniae af-E17 displayed higher activity (0.24 U/mg) followed by Klebsiella oxytoca af-G4 (0.20 U/mg). Conclusion: Habitat-specific innovations are being explored for novel compounds for therapeutic applications. This study throws a light on selection of carbohydrate degrading bacteria from a new source i.e., GUT of honeybee.

High Amylase Production by a Novel Strain of Bacillus amyloliquefaciens M37 Isolated from Can Gio Mangrove Forest, Vietnam

Amylases are one of the most important industrial enzymes and find applications in many areas such as textiles, chemicals, food, and pharmaceuticals. Most of the amylases are derived from microbes. The objective of the present study was to evaluate amylase production by a bacterium isolated from the Can Gio mangrove forest. The bacterium was identified as a species of genus Bacillus based on morphological and biochemical characteristics. The analysis of 16S rRNA sequences was then confirmed that this strain belonged to Bacillus amyloliquefaciens species (100% similarity). The effect of culture conditions such as temperature, pH, and carbon sources on amylase production through shake-flask culture was investigated. Maximum amylase activity of 904 IU/mL was obtained after 24 h of cultivation in LB medium containing 1% soluble starch at 35oC and pH 7.0. The highest enzyme activity of 1279 IU/mL was achieved in the bioreactor after 30 h of cultivation at optimum conditions. In addition, B. amyloliquefaciens M37 can grow on soybean meal medium. The high bacterial cell number of 456 × 109 CFU/g and amylase activity of 1039 IU/g were obtained after 36 h of cultivation. This newly isolated B. amyloliquefaciens M37 could be a potential producer for industrial amylase production and probiotics with commercial implications.