Search results for: Enzymatic

Authors: Rotchanaphan Hengaroonprasan, Malinee Sriariyanun, Prapakorn Tantayotai, Supacharee Roddecha, Kraipat Cheenkachorn

Abstract:

Lignocellolusic material is a substance that is resistant to be degraded by microorganisms or hydrolysis enzymes. To be used as materials for biofuel production, it needs pretreatment process to improve efficiency of hydrolysis. In this work, chemical pretreatments on rice straw using three diluted organic acids, including acetic acid, citric acid, oxalic acid, were optimized. Using Response Surface Methodology (RSM), the effect of three pretreatment parameters, acid concentration, treatment time, and reaction temperature, on pretreatment efficiency were statistically evaluated. The results indicated that dilute oxalic acid pretreatment led to the highest enhancement of enzymatic saccharification by commercial cellulase and yielded sugar up to 10.67 mg/ml when using 5.04% oxalic acid at 137.11 oC for 30.01 min. Compared to other acid pretreatment by acetic acid, citric acid, and hydrochloric acid, the maximum sugar yields are 7.07, 6.30, and 8.53 mg/ml, respectively. Here, it was demonstrated that organic acids can be used for pretreatment of lignocellulosic materials to enhance of hydrolysis process, which could be integrated to other applications for various biorefinery processes. 

Keywords: Lignocellolusic biomass, pretreatment, organic acid response surface methodology, biorefinery.

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Authors: Feng Chia Hsieh, Sheng Kuo Hsieh, Tzyy Rong Jinn

Abstract:

Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).

Keywords: Dipeptidyl-Peptidase IV, Phenylmethylsulfonyl fluoride; Serine protease, Sitagliptin, Vespa basalis

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Authors: Michele R. Spier, Denise N. X. Salmon, Renato L. Binati, Luíza C. Piva, Adriane B.P. Medeiros, Carlos R. Soccol

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Phytases are enzymes used as an important component in monogastric animals feeds in order to improve phosphorous availability, since it is not readily assimilated by these animals in the form of the phytate presented in plants and grains. As these enzymes are used in industrial activities, they must retain its catalytic activities during a certain storage period. This study presents information about the stability of 4 different phytases, produced by four macromycetes fungi through solid-state fermentation (SSF). There is a lack of data in literature concerning phytase from macromycetes shelf-life in storage conditions at room, cooling and freezing temperatures. The 4 phytases from macromycetes still had enzymatic activities around 100 days of storage at room temperature. At cooling temperature in 146 days of studies, the phytase from G. stipitatum was the most stable with 44% of the initial activity, in U.gds (units per gram of dried fermented substrate). The freezing temperature was the best condition storage for phytases from G. stipitatum and T. versicolor. Each condition provided a study for each mushroom phytase, totalizing 12 studies. The phytases showed to be stable for a long period without the addition of additives.

Keywords: macromycetes, phytase, solid-state fermentation, wheat bran, stability

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Authors: Aditi Shukla, Ambarish S. Vidyarthi, Subir Samanta

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The worldwide prevalence of H3N2 influenza virus and its increasing resistance to the existing drugs necessitates for the development of an improved/better targeting anti-influenza drug. H3N2 influenza neuraminidase is one of the two membrane-bound proteins belonging to group-2 neuraminidases. It acts as key player involved in viral pathogenicity and hence, is an important target of anti-influenza drugs. Oseltamivir is one of the potent drugs targeting this neuraminidase. In the present work, we have taken subtype N2 neuraminidase as the receptor and probable analogs of oseltamivir as drug molecules to study the protein-drug interaction in anticipation of finding efficient modified candidate compound. Oseltamivir analogs were made by modifying the functional groups using Marvin Sketch software and were docked using Schrodinger-s Glide. Oseltamivir analog 10 was detected to have significant energy value (16% less compared to Oseltamivir) and could be the probable lead molecule. It infers that some of the modified compounds can interact in a novel manner with increased hydrogen bonding at the active site of neuraminidase and it might be better than the original drug. Further work can be carried out such as enzymatic inhibition studies; synthesis and crystallizing the drug-target complex to analyze the interactions biologically.

Keywords: H3N2 Influenza, Neuraminidase, Oseltamiviranalogs, structure based drug designing

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Authors: Nasim Shaibani, Saba Ghazvini, Mohammad R. Andalibi, Soheila Yaghmaei

Abstract:

Nowadays there is a growing interest in biofuel production in most countries because of the increasing concerns about hydrocarbon fuel shortage and global climate changes, also for enhancing agricultural economy and producing local needs for transportation fuel. Ethanol can be produced from biomass by the hydrolysis and sugar fermentation processes. In this study ethanol was produced without using expensive commercial enzymes from sugarcane bagasse. Alkali pretreatment was used to prepare biomass before enzymatic hydrolysis. The comparison between NaOH, KOH and Ca(OH)2 shows NaOH is more effective on bagasse. The required enzymes for biomass hydrolysis were produced from sugarcane solid state fermentation via two fungi: Trichoderma longibrachiatum and Aspergillus niger. The results show that the produced enzyme solution via A. niger has functioned better than T. longibrachiatum. Ethanol was produced by simultaneous saccharification and fermentation (SSF) with crude enzyme solution from T. longibrachiatum and Saccharomyces cerevisiae yeast. To evaluate this procedure, SSF of pretreated bagasse was also done using Celluclast 1.5L by Novozymes. The yield of ethanol production by commercial enzyme and produced enzyme solution via T. longibrachiatum was 81% and 50% respectively.

Keywords: Alkali pretreatment, bioethanol, cellulase, simultaneous saccharification and fermentation, solid statefermentation, sugarcane bagasse

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Authors: T. H. Derdzyan, K. A. Ghazaryan, G. A. Gevorgyan

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Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoesterase and acetate-esterase enzyme activities in the soils under the impact of metallurgical industrial activity in Lori marz (district) were investigated. The results of the study showed that the activities of the investigated enzymes in the soils decreased with increasing distance from the Shamlugh copper mine, the Chochkan tailings storage facility and the ore transportation road. Statistical analysis revealed that the activities of the enzymes were positively correlated (significant) to each other according to the observation sites which indicated that enzyme activities were affected by the same anthropogenic factor. The investigations showed that the soils were polluted with heavy metals (Cu, Pb, As, Co, Ni, Zn) due to copper mining activity in this territory. The results of Pearson correlation analysis revealed a significant negative correlation between heavy metal pollution degree (Nemerow integrated pollution index) and soil enzyme activity. All of this indicated that copper mining activity in this territory causing the heavy metal pollution of the soils resulted in the inhabitation of the activities of the enzymes which are considered as biological catalysts to decompose organic materials and facilitate the cycling of nutrients.

Keywords: Armenia, metallurgical industrial activity, heavy metal pollutionl, soil enzyme activity.

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Authors: F. Benkaci-Ali, R. Mékaoui, G. Scholl, G. Eppe

Abstract:

The new methods as accelerated steam distillation assisted by microwave (ASDAM) is a combination of microwave heating and steam distillation, performed at atmospheric pressure at very short extraction time. Isolation and concentration of volatile compounds are performed by a single stage. (ASDAM) has been compared with (ASDAM) with cryogrinding of seeds (CG) and a conventional technique, hydrodistillation assisted by microwave (HDAM), hydro-distillation (HD) for the extraction of essential oil from aromatic herb as caraway and cumin seeds. The essential oils extracted by (ASDAM) for 1 min were quantitatively (yield) and qualitatively (aromatic profile) no similar to those obtained by ASDAM-CG (1 min) and HD (for 3 h). The accelerated microwave extraction with cryogrinding inhibits numerous enzymatic reactions as hydrolysis of oils. Microwave radiations constitute the adequate mean for the extraction operations from the yields and high content in major component majority point view, and allow to minimise considerably the energy consumption, but especially heating time too, which is one of essential parameters of artifacts formation. The ASDAM and ASDAM-CG are green techniques and yields an essential oil with higher amounts of more valuable oxygenated compounds comparable to the biosynthesis compounds, and allows substantial savings of costs, in terms of time, energy and plant material.

Keywords: Microwave, steam distillation, caraway, cumin, cryogrinding, GC-MS, GCxGC-MS.

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Authors: Ishtiaq Ahmed, Muhammad Naeem, Abdul Shakoor, Zaheer Ahmed, Hafiz Muhammad Nasir Iqbal

Abstract:

The present study was conducted to observe the effect of Plantago psyllium on blood glucose and cholesterol levels in normal and alloxan induced diabetic rats. To investigate the effect of Plantago psyllium 40 rats were included in this study divided into four groups of ten rats in each group. One group A was normal, second group B was diabetic, third group C was non diabetic and hypercholesterolemic and fourth group D was diabetic and hypercholesterolemic. Two groups B and D were made diabetic by intraperitonial injection of alloxan dissolved in 1mL distilled water at a dose of 125mg/Kg of body weight. Two groups C and D were made hypercholesterolemic by oral administration of powder cholesterol (1g/Kg of body weight). The blood samples from all the rats were collected from coccygial vein on 1st day, then on 21st and 42nd day respectively. All the samples were analyzed for blood glucose and cholesterol level by using enzymatic kits. The blood glucose and cholesterol levels of treated groups of rats showed significant reduction after 7 weeks of treatment with Plantago psyllium. By statistical analysis of results it was found that Plantago psyllium has anti-diabetic and hypocholesterolemic activity in diabetic and hypercholesterolemic albino rats.

Keywords: Albino rats, alloxan, Plantago psyllium, statistical analysis

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Authors: A. Sharma, R. Tewari, S. K. Soni

Abstract:

Biofuels production has come forth as a future technology to combat the problem of depleting fossil fuels. Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient method and catching the eye of scientific community. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus main objective of the present study was to optimize composition of medium components for enhancing cellulase production by newly isolated strain of Bacillus tequilensis. Nineteen factors were taken into account using statistical Plackett-Burman Design. The significant variables influencing the cellulose production were further employed in statistical Response Surface Methodology using Central Composite Design for maximizing cellulase production. The optimum medium composition for cellulase production was: peptone (4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L), Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride (0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at 37oC. Analysis of variance (ANOVA) revealed high coefficient of determination (R2) of 0.99. Maximum cellulase productivity of 11.5 IU/ml was observed against the model predicted value of 13 IU/ml. This was found to be optimally active at 60oC and pH 5.5.

Keywords: Bacillus tequilensis, CMCase, Submerged Fermentation, Optimization, Plackett-Burman Design, Response Surface Methodology.

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Authors: Vasudha Bansal, M. L. Singla, C. Ghanshyam

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The effect of nonthermal pulsed electric field (PEF) and thermal treatment (90⁰C for 60s) was studied on quality parameters of emblica officinalis juice for the period of 6 weeks at 4⁰C using monopolar rectangular pulse of 1µs width. The PEF treatment was given using static chamber at 24kV/cm for 500µs. The quality of emblica officinalis juice was investigated in terms of non enzymatic browning index (NEBI), 5-hydroxymethyl-2-furfural (HMF), total polyphenol content and antioxidant capacity. ⁰Brix, pH and conductivity were evaluated as physical parameters. The aim of the work was to investigate the effect of PEF on the retention of bioactive compounds and retardation of browning activity. The results showed that conventional thermal treatment had led to a significant (p < 0.05) decrease of 48.15% in polyphenol content (129.56 mg of GAE L^-1), with higher NEBI and HMF formation (p < 0.05) whilst PEF suppressed NEBI and retained higher polyphenol compounds (168.59 mg GAE L^-1) with limiting the loss to 32.56% along maximum free radical scavenging activity (92.07%). However, pH, ⁰brix and electrical conductivity of treated juice samples remain unaffected. Therefore, PEF can be considered as an effective nonthermal treatment for retaining bioactive compounds along suppressing browning of emblica juice.

 

Keywords: Emblica officinalis juice, Free radical scavenging activity, Pulsed electric field, Total polyphenol content.

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Authors: G. Khatisashvili, M. Gordeziani, G. Adamia, E. Kvesitadze, T. Sadunishvili, G. Kvesitadze

Abstract:

The ability of agricultural and decorative plants to absorb and detoxify TNT and RDX has been studied. All tested 8 plants, grown hydroponically, were able to absorb these explosives from water solutions: Alfalfa > Soybean > Chickpea> Chikling vetch >Ryegrass > Mung bean> China bean > Maize. Differently from TNT, RDX did not exhibit negative influence on seed germination and plant growth. Moreover, some plants, exposed to RDX containing solution were increased in their biomass by 20%. Study of the fate of absorbed [1-14ðí]-TNT revealed the label distribution in low and high-molecular mass compounds, both in roots and above ground parts of plants, prevailing in the later. Content of 14ðí in lowmolecular compounds in plant roots are much higher than in above ground parts. On the contrary, high-molecular compounds are more intensively labeled in aboveground parts of soybean. Most part (up to 70%) of metabolites of TNT, formed either by enzymatic reduction or oxidation, is found in high molecular insoluble conjugates. Activation of enzymes, responsible for reduction, oxidation and conjugation of TNT, such as nitroreductase, peroxidase, phenoloxidase and glutathione S-transferase has been demonstrated. Among these enzymes, only nitroreductase was shown to be induced in alfalfa, exposed to RDX. The increase in malate dehydrogenase activities in plants, exposed to both explosives, indicates intensification of Tricarboxylic Acid Cycle, that generates reduced equivalents of NAD(P)H, necessary for functioning of the nitroreductase. The hypothetic scheme of TNT metabolism in plants is proposed.

Keywords: Higher plants, TNT, RDX, transformation.

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Authors: Sharareh Mohseni, Reihaneh Ahmadzadeh Ghavidel

Abstract:

Tomato powder has good potential as substitute of tomato paste and other tomato products. In order to protect physicochemical properties and nutritional quality of tomato during dehydration process, investigation was carried out using different drying methods and pretreatments. Solar drier and continuous conveyor (tunnel) drier were used for dehydration where as calcium chloride (CaCl2), potassium metabisulphite (KMS), calcium chloride and potassium metabisulphite (CaCl2 +KMS), and sodium chloride (NaCl) selected for treatment.. lycopene content, dehydration ratio, rehydration ratio and non-enzymatic browning in addition to moisture, sugar and titrable acidity were studied. Results show that pre-treatment with CaCl2 and NaCl increased water removal and moisture mobility in tomato slices during drying of tomatoes. Where CaCl2 used along with KMS the NEB was recorded the least compared to other treatments and the best results were obtained while using the two chemicals in combination form. Storage studies in LDPE polymeric and metalized polyesters films showed less changes in the products packed in metallized polyester pouches and even after 6 months lycopene content did not decrease more than 20% as compared to the control sample and provide extension of shelf life in acceptable condition for 6 months. In most of the quality characteristics tunnel drier samples presented better values in comparison to solar drier.

Keywords: Dehydration, Tomato powder, Lycopene, Browning

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Authors: Reihaneh Ahmadzadeh Ghavidel, Mehdi Ghiafeh Davoodi

Abstract:

Tomato powder has good potential as substitute of tomato paste and other tomato products. In order to protect physicochemical properties and nutritional quality of tomato during dehydration process, investigation was carried out using different drying methods and pretreatments. Solar drier and continuous conveyor (tunnel) drier were used for dehydration where as calcium chloride (CaCl2), potassium metabisulphite (KMS), calcium chloride and potassium metabisulphite (CaCl2 +KMS), and sodium chloride (NaCl) selected for treatment.. lycopene content, dehydration ratio, rehydration ratio and non-enzymatic browning in addition to moisture, sugar and titrable acidity were studied. Results show that pre-treatment with CaCl2 and NaCl increased water removal and moisture mobility in tomato slices during drying of tomatoes. Where CaCl2 used along with KMS the NEB was recorded the least compared to other treatments and the best results were obtained while using the two chemicals in combination form. Storage studies in LDPE polymeric and metalized polyesters films showed less changes in the products packed in metallized polyester pouches and even after 6 months lycopene content did not decrease more than 20% as compared to the control sample and provide extension of shelf life in acceptable condition for 6 months. In most of the quality characteristics tunnel drier samples presented better values in comparison to solar drier.

Keywords: Dehydration, Tomato powder, Lycopene, Browning

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Authors: Senem Suna, Canan Ece Tamer, Ömer Utku Çopur

Abstract:

In this research, dried linden (Tilia argentea) leaves and blossoms were used as a raw material for mineral enriched herbal tea beverage production. For this aim, %1 dried linden was infused with boiling water (100 °C) for 5 minutes. After cooling, sucrose, citric acid, ascorbic acid, natural lemon flavor and natural mineral water were added. Beverage samples were plate filtered, filled into 200-mL glass bottles, capped then pasteurized at 98 °C for 15 minutes. Water soluble dry matter, titratable acidity, ascorbic acid, pH, minerals (Fe, Ca, Mg, K, Na), color (L*, a*, b*), turbidity, bioaccessible phenolics and antioxidant capacity were analyzed. Water soluble dry matter, titratable acidity, and ascorbic were determined as 7.66±0.28 g/100 g, 0.13±0.00 g/100 mL, and 19.42±0.62 mg/100 mL, respectively. pH was measured as 3.69. Fe, Ca, Mg, K and Na contents of the beverage were determined as 0.12±0.00, 115.48±0.05, 34.72±0.14, 48.67±0.43 and 85.72±1.01 mg/L, respectively. Color was measured as 13.63±0.05, -4.33±0.05, and 3.06±0.05 for L*, a*, and b* values. Turbidity was determined as 0.69±0.07 NTU. Bioaccessible phenolics were determined as 312.82±5.91 mg GAE/100 mL. Antioxidant capacities of chemical (MetOH:H₂O:HCl) and physiological extracts (in vitro digestive enzymatic extraction) with DPPH (27.59±0.53 and 0.17±0.02 μmol trolox/mL), FRAP (21.01±0.97 and 13.27±0.19 μmol trolox/mL) and CUPRAC (44.71±9.42 and 2.80±0.64 μmol trolox/mL) methods were also evaluated. As a result, enrichment with natural mineral water was proposed for the development of functional and nutritional values together with a good potential for commercialization.

Keywords: Antioxidant capacity, bioaccessibility, herbal tea beverage, linden.

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Authors: Mohammad Asif

Abstract:

A large number of chemical, bio-chemical and pollution-control processes use heterogeneous fixed-bed reactors. The use of finite hollow cylindrical catalyst pellets can enhance conversion levels in such reactors. The absence of the pellet core can significantly lower the diffusional resistance associated with the solid phase. This leads to a better utilization of the catalytic material, which is reflected in the higher values for the effectiveness factor, leading ultimately to an enhanced conversion level in the reactor. It is however important to develop a rigorous heterogeneous model for the reactor incorporating the two-dimensional feature of the solid phase owing to the presence of the finite hollow cylindrical catalyst pellet. Presently, heterogeneous models reported in the literature invariably employ one-dimension solid phase models meant for spherical catalyst pellets. The objective of the paper is to present a rigorous model of the fixed-bed reactors containing finite hollow cylindrical catalyst pellets. The reaction kinetics considered here is the widely used Michaelis–Menten kinetics for the liquid-phase bio-chemical reactions. The reaction parameters used here are for the enzymatic degradation of urea. Results indicate that increasing the height to diameter ratio helps to improve the conversion level. On the other hand, decreasing the thickness is apparently not as effective. This could however be explained in terms of the higher void fraction of the bed that causes a smaller amount of the solid phase to be packed in the fixed-bed bio-chemical reactor.

Keywords: Fixed-bed reactor, Finite hollow cylinder, Catalyst pellet, Conversion, Michaelis-Menten kinetics.

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Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA₄₄₀₄. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: Recombinant tissue plasminogen activator, plant cell comportment, leader signals, transgenic tobacco.

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Authors: Aka S. Koffi, N’Goran Yao, Philippe Bastide, Denis Bruneau, Diby Kadjo

Abstract:

Cocoa beans (Theobroma cocoa L.) are the main components for chocolate manufacturing. The beans must be correctly fermented at first. Traditional process to perform the first fermentation (lactic fermentation) often consists in confining cacao beans using banana leaves or a fermentation basket, both of them leading to a poor product thermal insulation and to an inability to mix the product. Box fermenter reduces this loss by using a wood with large thickness (e>3cm), but mixing to homogenize the product is still hard to perform. Automatic fermenters are not rentable for most of producers. Heat (T>45°C) and acidity produced during the fermentation by microbiology activity of yeasts and bacteria are enabling the emergence of potential flavor and taste of future chocolate. In this study, a cylindro-rotative fermenter (FCR-V1) has been built and coconut fibers were used in its structure to confine heat. An axis of rotation (360°) has been integrated to facilitate the turning and homogenization of beans in the fermenter. This axis permits to put fermenter in a vertical position during the anaerobic alcoholic phase of fermentation, and horizontally during acetic phase to take advantage of the mid height filling. For circulation of air flow during turning in acetic phase, two woven rattan with grid have been made, one for the top and second for the bottom of the fermenter. In order to reduce air flow during acetic phase, two airtight covers are put on each grid cover. The efficiency of the turning by this kind of rotation, coupled with homogenization of the temperature, caused by the horizontal position in the acetic phase of the fermenter, contribute to having a good proportion of well-fermented beans (83.23%). In addition, beans’pH values ranged between 4.5 and 5.5. These values are ideal for enzymatic activity in the production of the aromatic compounds inside beans. The regularity of mass loss during all fermentation makes it possible to predict the drying surface corresponding to the amount being fermented.

Keywords: Cocoa fermentation, fermenter, microbial activity, temperature, turning.

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Authors: Mihriban Korukluoglu, Goksen Arik, Cagla Erdogan, Selen Kocakoglu

Abstract:

Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Keywords: Antagonistic effect, kefir, lactic acid bacteria (LAB), synergistic, yeast.

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