Extraction, Partial Purification and Characterization of Bromelain from Pineapple (Ananas Comosus) Crown, Core and Peel Waste

Abstract Ananas Comosus (also known as pineapple) is a part of Bromeliaceae family and it is consumed as food as well as folk medicine for the treatment of various diseases. It is reported that pineapple is a rich source of bromelain, a cysteine protease and it is considered as an important enzyme in different industries due to its significant therapeutic and industrial applications such as anticancer, anti-inflammatory and meat tenderizing. Bromelain is mostly present in fruit and stem of pineapple, but it is reported that crown, core, and peels, which constitute the waste of the pineapple plant, also contain bromelain but limited data is available. Therefore, the proposed study aimed at utilizing pineapple waste for the extraction and characterization of bromelain. Firstly, crude bromelain was extracted with phosphate buffer (pH 7), then it was subjected to partial purification using different fractions of ammonium sulphate (NH4)2SO4 such as 30, 40, 50 and 60% followed by desalting and concentration. Enzyme activity was calculated by using casein digesting unit (CDU) method. The results demonstrated that the crown bromelain showed highest purification of 4.34-fold at 30% (NH4)2SO4 saturation, whereas core and peel bromelain showed highest purification of 2.75 and 2.59-fold at 40% (NH4)2SO4 saturation. The molecular weight of crude and partially purified bromelain was determined by SDS-PAGE analysis and found to be 26 KDa. The pH and thermal stability of all the parts of pineapple showed maximum stability at pH 7 and at 35oC temperature.

Improved Cultural Conditions for Methionine Accumulation in Submerged Cultivation of Bacillus cereus S8.

Aims: To improve the cultural conditions for enhanced methionine production by Bacillus cereus S8 Study design: Study of the fermentation process in shake flask culture. Place and Duration of Study: Department of Applied Microbiology and Brewing, Nnamdi Azikwe University, Awka, Nigeria between 2011 to 2012. Methodology: The effects of medium/fermenter volume ratio, carbon and nitrogen sources, growth stimulators, vitamins and amino acid on methionine accumulation in the broth culture of Bacillus cereus S8 were investigated. The time course for methionine production was also studied. Results: A 20% medium/fermenter volume ratio improved methionine yield. Glucose and ammonium sulphate at 6.0 and 1.0% respectively stimulated methionine accumulation by Bacillus cereus S8. Yeast extract, peptone, DL-leucine and all vitamins studied enhanced methionine production. A methionine yield of 3.23mg/ml was produced after 96h fermentation and at a pH of 6.90. Conclusion: Improving the cultural conditions of Bacillus cereus S8 in submerged medium stimulated methionine increase.

Cryoprotection effectiveness of low concentrations of natural and lyophilized LDL (low density lipoproteins) on canine spermatozoa / Eficiência de baixas concentrações de lipoproteínas de baixa densidade natural e liofilizada na crioproteto de espermatozoides caninos

The aim of this study was to evaluate the use of low concentrations of natural and lyophilized low density lipoprotein (LDL) from hen's egg yolk for cryopreservation of canine semen. Different ammonium sulphate concentrations were tested to extract LDL from egg yolk. The yolk was centrifuged, and LDL was isolated using 10, 20, 40, 45, or 50 percent ammonium sulphate solution (ASS). The LDL-rich floating fraction was collected for chemical characterization. Dry matter content was lowest (P<0.05) in the LDL extracted with the 50 percent ASS. The purification of LDL increased in association with increasing ammonium sulphate concentrations. SDS-PAGE showed that the 50 percent ASS solution yielded a purer fraction of LDL from egg yolk. For semen cryopreservation, TRIS extender was used replacing 20 percent egg yolk (control) by natural or lyophilized LDL using 1, 2, and 3 percent (w/v). Semen was centrifuged (755Xg for 7 min), diluted with one of the extenders, packed into 0.5mL straws (100x106 sperm/mL), and placed in a programmable cryopreservation machine. Thawed semen (37°C/ 30s) was analyzed for sperm motility, morphology, and by the hypoosmotic and epifluorescence tests (CFDA/ PI). Natural LDL extracted with 50 percent ASS was as effective as whole egg yolk to preserve canine frozen sperm when using low concentrations. The lyophilized LDL, mainly in the two higher concentrations tested (2 and 3 percent), was unsuitable to maintain the effectiveness of the LDL cryoprotective effect on dog sperm...

O objetivo deste estudo foi avaliar o uso de baixas concentrações da lipoproteína de baixa densidade (LBD) extraída da gema do ovo de galinha, nas formas natural e liofilizada, na criopreservado do sêmen canino. Diferentes concentrações de sulfato de amônio também foram testadas na extrato da LBD da gema do ovo. A gema foi centrifugada, sendo a LBD isolada usando-se soluto saturada de sulfato de amônio (SSA) nas concentrações de 10, 20, 40, 45 e 50 por cento. A frado rica em LBD foi coletada para caracterizado química. O conteœdo de matéria seca foi menor (P<0,05) na LBD extraída com SSA 50 por cento. A pureza da LBD melhorou medida que se aumentou a concentrado de SSA utilizada. SDS-PAGE mostrou que a SSA 50 por cento produziu uma frado mais pura de LBD oriunda da gema do ovo. Para o congelamento de sêmen, o meio diluidor TRIS teve a gema do ovo a 20 por cento (controle) substituída pela LBD a 1, 2 e 3 por cento (p/v), nas formas natural e liofilizada. O sêmen foi centrifugado (755xg por 7min), diluído em um dos meios diluidores em teste e envasado em palhetas de 0,5mL (100x106 sptz/mL), sendo congelado em máquina de congelamento programável. O sêmen descongelado (37°C/30s) foi analisado quanto motilidade e morfologia espermática e nos testes hiposm-tico e de epifluorescência (DACF/IP). A LBD natural extraída com SSA 50 por cento foi tão eficiente quanto a gema do ovo na preservado do espermatozoide canino congelado nas baixas concentrações testadas. A LBD liofilizada, principalmente as duas maiores concentrações (2 e 3 por cento), não foi adequada para manter o efeito crioprotetor da LBD sobre o espermatozoide canino...

Characterization and Purification of Protease enzyme.

Protein is one of the three major food groups needed for proper nutrition. Proteolytic enzymes or proteinases are the group of enzymes whose catalytic function is to hydrolyze (breakdown) proteins. Production and partial purification of protease enzyme by Bacillus subtilis was the aim of this study. Bacillus subtilis was allowed to grow in shake flask broth culture, 3.5L and 7L fermenters for purpose of inducing protease enzyme. We are finding out the effect of minerals which are useful for the production of protease by Plackett Burman design. Minerals such as magnesium sulphate, potassium dihydrogen phosphate and manganese sulphate were showing the results for the production of protease. The protease enzyme was purified by ultra filtration, ammonium sulphate precipitation, dialysis, and lyophilization.The activity of protease was increased as there was increase in the enzyme concentration. Purified protease enzyme had a maximum activity at pH 9.0 of carbonate buffer and the optimum incubation time was 48hr. The protease assay is done for the crude enzyme at different temperature. It showed greater activity at 50 °C but after that it started decreasing the activity so, we had selected the temperature at 40 ℃ for the good activity..

Fontes e doses de nitrogênio em cobertura no feijoeiro de inverno irrigado no sistema plantio direto / Sources and doses of sidedressing nitrogen on irrigated common bean cropped in winter in no tillage system

O feijoeiro é uma leguminosa de grande importância na economia brasileira, e o nitrogênio é o nutriente absorvido em maior quantidade. O manejo da adubação nitrogenada é de extrema importância no sentido de oferecer maior viabilidade econômica, além de aumentar a eficiência da planta na utilização dos recursos disponíveis. O objetivo do trabalho foi estudar o efeito de fontes e doses de nitrogênio em cobertura no desenvolvimento e produtividade do feijoeiro de inverno no sistema plantio direto, correlacionado com uma análise econômica simples. O delineamento experimental foi de blocos casualizados em esquema fatorial 3x3, constituídos pela combinação de três fontes de nitrogênio (sulfato de amônio, uréia e mistura - sulfato de amônio ½ do N + uréia ½ do N) e diferentes doses de nitrogênio em cobertura (zero, 40 e 80 kg ha-1, aplicado na fase V4-3), com quatro repetições. O projeto foi conduzido no município de Selvíria (MS), no período de outono-inverno de 2004. O solo do local é um LATOSSOLO VERMELHO Distrófico argiloso. Conclui-se que independente da fonte de N, o aumento da adubação nitrogenada proporciona incremento na produtividade do feijoeiro de inverno até a dose de 80 kg ha-1, sendo que esta proporciona em média, aumento de 25 % na produtividade comparado com a testemunha (sem N em cobertura). A uréia é a fonte de nitrogênio de maior eficiência econômica.

The common bean is a leguminous of great importance in the Brazilian economy and nitrogen is the taken up nutrient in larger amount. Nitrogen fertilization management is of extreme importance to offer larger economical viability, besides increasing the efficiency of plant in the use of the available resources. The objective this study was to evaluate the effect of sources and doses of sidedressing nitrogen in the development and yield of winter common bean in no tillage system, as well as evaluate its economical viability. A randomized blocks design was used, in a factorial scheme 3x3 with 9 treatments constituted by three sources of nitrogen (ammonium sulphate, urea and ammonium sulphate ½ of N + urea ½ of N, applied at V4-3 stadium) and different doses of sidedressing nitrogen (0, 40, 80 kg ha-1) in four replications. The study was conducted in Selvíria county, MS State in 2004 in no season crop period, in a dystrophic Haplustox soil. The conclusion: independent of nitrogen source, nitrogen fertilization increasing provides increment in yield of winter common bean up to dose 80 kg/ha, and this provides, on average, an increase of 20% in yield compared with control (without sidedressing nitrogen). The urea is the nitrogen source of larger economical efficiency

Doses e fontes de nitrogênio na recuperação de pastagens do capim-marandu / Nitrogen doses and sources on pasture recuperation of grass marandu

Avaliaram-se o efeito de doses e fontes de nitrogênio na recuperação do capim-marandu, por um período de três anos, em pastagem estabelecida há mais de 10 anos, com baixa produção de forragem. O delineamento experimental foi em blocos completos ao acaso, com três repetições. Nas parcelas foi utilizado o fatorial 2 x 4, sendo duas fontes de nitrogênio (sulfato de amônio e uréia) e quatro doses de nitrogênio (0, 100, 200 e 300kg ha-1 ano-1). Nas subparcelas, foram alocados os três anos (2004, 2005 e 2006), referentes ao tempo de recuperação da pastagem. A aplicação de nitrogênio foi determinante para a recuperação do capim-marandu. A maior produção de massa seca foi observada no segundo ano e o maior teor de proteína bruta no terceiro ano de recuperação da pastagem. As maiores doses de nitrogênio promoveram acréscimos lineares na produção de massa seca e redução nos teores de fibra em detergente neutro e fibra em detergente ácido. O sulfato de amônio promoveu maior produção de massa seca do que a ureia, em todas as doses e anos avaliados.

The effects of nitrogen doses and sources were evaluated on pasture recuperation of grass marandu, in a three-year period. The pasture was established for more than ten years and it was presenting low herbage production being considered in moderate degradation phase. The experiment was arranged in a randomized complete block design with split-plots and three replications, in a 2x4 factorial, being two sources of N (ammonium sulphate and urea) and four doses of N (0, 100, 200, and 300kg ha-1 yr-1). The time of pasture evaluation was represented by the years 2004, 2005, and 2006. The highest dry matter production was observed in the second year and the highest crude protein in the third one. The highest nitrogen doses promoted linear increase on dry mass production and decrease in fiber concentration and in neutral and acid detergents. Ammonium sulphate resulted in higher dry mass production than urea in all doses applied and evaluated years.

Effect of Different Media on MPN Estimation of Ammonia-oxidizing Bacteria in Lakes / 微生物学通报

The MPN method was used to enumerate ammonia-oxidizing bacteria (AOB) in water and sediments of several shallow lakes. The suitable incubation time, medium types and substrate (ammonium sulphate) concentrations were studied. The results showed that, MPN values increased with the incubation time, reaching a stable maximum at some time stages, which was 40 days in all the samples for MSF medium. Among the three media used (XZ-AOB、MSF、SW), MSF give the highest MPN value. In addition, am- monium sulphate concentration in medium was an important factor affecting MPN estimation of AOB. Compared to AOB in lake sediments, AOB in lake water was more sensitive to ammonium sulphate concentration.

Ammonium sulphate induced stress related alterations in the respiratory epithelium of the airbreathing organ of the catfish Heteropneustes fossilis (Bloch).

In this paper, histopathological changes in the inner lining of the accessory respiratory organ of Heteropneustes fossilis following exposure to sublethal concentration (0·2 g I -1) of ammonium sulphate (3 mg I-1 total ammonia-N) has been described. The goblet cells show periodic increased followed by decreased secretory activities. Necrosis and shedding of the epithelial cells over the secondary lamellae cause periodic haemorrhages which lead to degeneration and decreased number of secondary lamellae. Subsequently regeneration takes place each time as evidenced by the appearance of inflammatory tissue. Fusion of more than one secondary lamellae is also common. Regeneration also leads to uncontrolled hyperplasia of haphazardly arranged epithelial cells. This hyperplasia causes increased distance of respiratory blood-air barrier in the secondary lamellae, leading to impaired normal aerial respiration.