Sensory evaluation and contamination test on mango fruit (Mangifera indica) puree

Cancer is a disease which is known Ihroughoul the world. There arc many available commercial and alternalive treatments to Ireat cancer. Mango (Mungiferu indica L) is one the popular trOPical fru;ts found in Malaysia. A wide variety of mang0C5 can easily be fouod in this country, Mangoes are known for the;r vibrant flesh colour. juicy texture. sweet flavour. vitamins and minerals. Besides that. mango also contains anticancer and antiproliferation properties. Siamese mango was sekcted for oplimilation of purec sterilizatiOtl. Mango puree was Sterilized and stored for approximately five days. The sensory evaluation and contamination test was performed on sterilized lmd stored puree, Contamination test shows no growth for Staphylococcus aure"s and E. coli/colifonn. Puree 2 (8S°C. IS min sterilization lime and lC!u 970) and Puree 10 (8)<C, 8 min sterilization time. and IC~ 640) ha\'e satisfied most respondents, Variation of temperature and time for puree sterilization does affect the appearance. taste and tCJlture oftl>c puree

Case Study: Recombinant Bromelain Downstream Processing

This chapter presents details on the purification, formulation and drying of recombinant bromelain. The wide range of applications of recombinant bromelain has increased interest in finding viable purification techniques for large-scale production. An affinity chromatography technique was developed by Amid and co-workers (Expression, purification, and characterization of a recombinant stem bromelain from Ananas comosus, Process Biochem 46:2232–2239, 2011) to purify recombinant bromelain. However, this technique presented low recovery and small sample loading capacity and thus is not suitable as a purification tool in the largescale production of recombinant bromelain. An aqueous two-phase system is one alternative method that we use to purify recombinant bromelain, as it reliable and easy to scale up and has a low cost. As part of avoiding cysteine degradation, spray drying the purified recombinant protein with maltodextrin as an excipient provides the possibility of preserving its activity and creating fine particles that are suitable for end-product application. The processes of the purification, formulation and spray drying of recombinant bromelain are explained briefly

Screening and evaluation of anticancer property in mango fruit mangifera indica

Cancer is a disease which known throughout the world. There are many available commercial and alternative treatments to treat cancer. Mango (Mangi(era indica L) is one the popular tropical fruits found in Malaysia. Mango is a wide variety that can easily be found in this country. It is known for its vibrant flesh colour, juicy texture, sweet flavour, vitamins and minerals. This study aimed to evaluate the anticancer property of mango variety that available in Malaysia (Siamese, Honey, Kuinin and Chokonan) towards human breast cancer cell (MCF-7). Based on the result. it turned out that Kuinin has the lowest ICso value. 300 llgmL-1 compared to other mango variety. suggesting that Kuinin as fruit has potential in preventing breast cancer

Optimization of heat sterilization on mango fruit (mangifera indica) puree and effects towards cancer treatment

Cancer is a disease Which known throughout the world. There are many available commercial and alternative treatments to treat cancer. Mango (Mangifera indica L.) is one the popular tropical fruits found in Malaysia. Mango is a wide variety that can easily be found in this country. It is known for its vibrant flesh colour, juicy texture, sweet flavour, vitamins and minerals. Besides that, mango also contains anticancer and antiprollferation properties. Siamese mango was selected for optimization of puree sterilization. The Design Expert software version 6.0.8 was used to design the experiment to optimize puree sterilization conducted using Central Composite Design with to factor which are variation of temperature (85°C-95°C) and time (1-16 minutes). The optimum condition for the sterilization treatment was at 90°C and 30 seconds holding time

Common Laboratory Procedure

This chapter presents all common laboratory procedures or protocols that researchers normally perform during studies of recombinant enzyme production. The principle and theory behind each experiment is presented to help the reader understand each step that is conducted in an experiment so the reader will be able to select and modify the procedures according to the type of enzyme they are working with. Examples of procedures and the results obtained during recombinant bromelain production are used to develop readers’ understandin

Optimization on cell disruption of E. coli BL21-AI expressing recombinant bromelain

Recent advancements in recombinant DNA technology proved to be a promising and effective approach for more sustainable large scale productions of many therapeutic proteins. Nevertheless, since this approach involves expression of proteins in a nonnative host microorganism, the overall production processes are not straight-forward due to several common challenges, such as protein degradation, especially during cell disruption stage. As the process has been subjected to both protein and host-specific, a systematic process conditioning for maximal production of recombinant protein is therefore required. In this study, a simple approach to determine optimal conditions for cell disruption using ultrasonication to isolate recombinant bromelain from E. coli BL21- AI is reported. Suspension cells were lysed using ultrasonication which transmit sound wave in other to break the cell wall. An optimized condition was obtained by response surface methodology (RSM). A three factor face-centered central composite design (FCCD) was applied to obtain the optimal process conditions consisting of amplitude, cycle and bursting period. The prediction model was further validated. Therefore, under the optimal conditions, having 20% amplitude, 0.5s cycle, and 1 minute bursting period in three times process, the specific enzyme activity of the recombinan

Biosynthesis of nanoparticles using recombinant bromelain

Green chemical method was applied to synthesize nanoparticles using recombinant bromelain. Among the numerous applications of recombinant bromelain, there is still no research on nanoparticles synthesis which encourages its utilization in this study. Four chemicals which are copper (II) chloride dihydrate (CuCl2.2H2O), cerium nitrate hexahydrate (Ce(NO3)3.6H2O), sodium selenite (Na2SeO3), and iron (III) chloride hexahydrate (FeCl3.6H2O) were selected to be screened for the suitability in nanoparticles biosynthesis by recombinant bromelain. The nanoparticles formed were characterized by using UV-visible absorption spectra. The biosynthesis process then was optimized by varying the centrifugation speed, temperature, and time to get the maximum absorption and weight of nanoparticles through central composite design (CCD) tool. Only CuCl2.2H2O showed a positive result for the screening process which was represented by the formation of colloidal solution and a maximum absorption at 580 nm. Thus, optimization was carried out for this chemical. Based on the optimization model, maximum absorption and weight were predicted at 67.5°C, 2 hrs, and 9,600 rpm. These optimal conditions were validated by repeating the biosynthesis process. The absorption and weight of the nanoparticles depended on the reaction of the chemical with recombinant bromelain. 3D plots showed that the optimal condition for high responses mostly depends on temperature and time

Comparison of different cell disruption methods and cell extractant buffers for recombinant bromelain expressed in E.coli BL21-A1

The often-encountered problem such as protein degradation has driven various methods of cell lysis in obtaining recombinant protein post fermentation. In this paper, we compare methods such as homogenization, sonication, sonication with lysozyme and chemical lysis using B-PER reagent with lysozyme to extract the recombinant bromelain from E. coli BL21-AI. The sonication process is found to be the most effective in releasing recombinant bromelain without any pre-treatment. To obtain the high quality of protein from sonication method, the influence of different extractant buffer was investigated including phosphate buffer saline (PBS), PBS containing cysteine and EDTA (PBS-CE), and sodium phosphate buffer containing cysteine and EDTA (EB-CE). The highest specific enzyme activity was obtained when it was extracted with EB-CE buffer. Under sodium dodecyl sulfate polyacrylamide gel electrophoresis, the recombinant bromelain showed protein band at 55kDa. In conclusion, the sonication method with extractant buffer containing 100mM phosphate buffer pH7.0 with 15 mM cysteine and 2 mM EDTA (EB-CE) was shown to give high specific activity of recombinant bromelain

Comparison of Different Cell Disruption Methods And Cell Extractant Buffers For Recombinant Bromelain Expressed in E.COLI BL21-A1

The often-encountered problem such as protein degradation has driven various methods of cell lysis in obtaining recombinant protein post fermentation. In this paper, we compare methods such as homogenization, sonication, sonication with lysozyme and chemical lysis using B-PER reagent with lysozyme to extract the recombinant bromelain from E. coli BL21-AI. The sonication process is found to be the most effective in releasing recombinant bromelain without any pre-treatment. To obtain the high quality of protein from sonication method, the influence of different extractant buffer was investigated including phosphate buffer saline (PBS), PBS containing cysteine and EDTA (PBS-CE), and sodium phosphate buffer containing cysteine and EDTA (EB-CE). The highest specific enzyme activity was obtained when it was extracted with EB-CE buffer. Under sodium dodecyl sulfate polyacrylamide gel electrophoresis, the recombinant bromelain showed protein band at 55kDa. In conclusion, the sonication method with extractant buffer containing 100mM phosphate buffer pH7.0 with 15 mM cysteine and 2 mM EDTA (EB-CE) was shown to give high specific activity of recombinant bromelain

Effects of operating conditions in spray drying of recombinant bromelain

The aim of the present study is to investigate the effect of operating parameter of a Buchi B290 mini spray dryer towards specific activity of recBromelain. Using Response Surface Methodology (RSM), face centred central composite design (FCCCD) was tabulated for optimization. Optimization process was conducted by varying the inlet air temperature (°C), gas flow height (mm) and pump settings (%) of the laboratory Buchi B290 Mini Spray Dryer to obtained optimal spray drying operating processes recovering high recBromelain specific activity as response. Results showed that the optimized process parameter having 126 °C air inlet temperature, 42 mm gas flow height and 12 % feed pump setting resulted in 0.119 ± 0.003 U/mgprotein of spray dried recBromelain with R2 of 90.12% thus this model was found to be significance and reliable