Open Access Original Research Article

Optimization of Aqueous Enzymatic Extraction of Flat-Europe Hybrid Hazelnut (Corylus heterophylla Fisch. × C. avellana L.) Oil and Analysis of Its Components

Chunmao Lv, Na Liu, Xianjun Meng, Changying Lu, Xiao Li

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BJI/2017/31140

Aim: The research aimed to evaluate the technological factors of the composite enzymatic extraction of Flat-European hybrid hazelnut oil, and analyze the fatty acid composition, to lay the foundations for the industrialized production of this oil.

Study Design: Based on single factor tests, the best conditions for the aqueous enzymatic extraction of the oil determined by response surface analysis.

Place and Duration of Study: Food Science College, Shenyang Agricultural University between March 2014 and December 2015.

Methodology: The composition of Flat-European hybrid hazelnut was determined using GC-MS.

Results: Selection of suitable commercial enzymes is an important factor that can affect the extraction efficiency. Based on single factor tests, best conditions for the aqueous enzymatic extraction of the oil, determined by response surface analysis, were as follows: pH 6.13, enzymatic hydrolysis temperature of 59.9°C, dose of papain equal to 6% of seed weight, dose of α-amylase equal to 4% of seed weight, seed/water ratio of 1:5 (w/v), and enzyme hydrolysis time of 4.78 hours. By means of GC-MS analysis oleic acid (77.22%), linoleic acid (14.06%), linolenic acid (0.13%), palmitic acid (5.04%), stearic acid (2.35%), and arachidonic acid (0.44%) were identified as the major compounds of the flat-European hybrid hazelnut oil.

Conclusion: The mixture of papain (6% of seed weight) and α-amylase (4% of seed weight) was the best enzyme combination for the aqueous enzymatic extraction of flat-European hybrid hazelnut oil. The best technological parameters for the oil extraction process were: seed/water ratio of 1:5 (w/v), pH value of 6.13, reaction temperature of 59.90°C, and reaction time of 4.78 h. With these conditions, the oil yield was 82.36%, where unsaturated fatty acids were 92% of the total.

Open Access Original Research Article

Sensitivity, Approach and Awareness about Genetically Modified Crops among Undergraduate Students: An Online Survey

Pravin Sukhadeo Gaikar, Rishikesh Haribhau Autade, Ramesh Madhav Jadhav, Amol Ramesh Savant, Rakeshkumar Sheshrao Choudhary, Ashok Vasantrao Kharde

Biotechnology Journal International, Page 1-10
DOI: 10.9734/BJI/2017/31555

The Survey-based studies provide the guidelines and feedback for policy makers about adopted and newborn technology. Biotechnology is the new branch of applied science and is emerging as the separate branch of the study in Agriculture (Agri-Biotechnology), Pharmacy (Pharma-Biotech), Environment Science (Environment Biotechnology), medical (Medical Biotechnology) and Industry (Industrial Biotechnology). The products of the biotechnology in many fields have been accepted by human society except Agriculture. After ‘Green Revolution’ the major breakthrough in Agriculture was an introduction of genetically modified crops but also brought controversies with it. There is a tug of war between GM crop supporters and Anti-GM crop organizations which have spread many myths in the society about this neonatal technology. Perception, approach and awareness about genetically modified crops issues among undergraduate students get affected due to ever increasing debate on the ‘Digital and Broadcasting Media’ platforms. To catch these digital techno-savvy young undergraduate student’s perception the web-based online survey was done through predesigned Google form. Total 136 respondents participated in a web-based online survey and the percentage of male and female participants were 48 and 52, respectively. The education branch wise participants in the study were highest for Agri-biotech (26%) and least for Engineering (13%), while for agriculture, pharmacy and agribusiness management the participation percentage was 16, 22 & 23, respectively. This study concluded that the students were affirmative towards the use of technology but receive awareness about the GM crops issue through unreliable source i.e. internet (30%), newspapers (21%) and television (16%). There is need of further continuous self-sustaining and long lasting awareness campaign with the self-motivated support from scientists working on the GM technology.

Open Access Original Research Article

Characterization of Spent Bleaching Earth and Its Utilization for Improving Manure-based Biogas Production

Anselm P. Moshi

Biotechnology Journal International, Page 1-13
DOI: 10.9734/BJI/2017/31601

Aim of the Study: To utilize spent bleaching earth (SBE) to improve biogas production efficiency through co-digesting with manure and provide a practical way of disposing the SBE.

Design of the Study: The SBE was collected from two edible oil refineries in Dar-es salaam, Tanzania. Composition was analysed in triplicate to determine its suitability for biogas production. Thereafter, the SBE was mixed with manure at (%) 0, 10, 20, 30, 40, 50 and 100 for biogas production.

Methodology: The residual oil from the SBE was recovered via solvent extraction system (soxtherm). Total solids, volatile solids and pH were determined using standard methods.  Physicochemical parameters of the SBE were determined using AOCS Official methods. Total carbon and total nitrogen were analysed using elemental analyser whereas the fatty acid profile of the residue oil was established using gas-chromatography techniques. Methane potential of mixtures SBE and manure at different proportions was investigated using a multi-channel analyser, methane potential test system in 500 mL glass bottles with reaction volume of 300 mL.

Place and Duration of Study: The study was completed in 2 years from 2015-2016. Samples of SBE were collected from two edible oil refineries in Dar-es salaam, Tanzania from March to July 2015 and all experiements were conducted between January and April 2016 at Tanzania Industrial Research and Development Organization.

Results: Two types of waste; soapstock ca.3150 metric tons and SBE ca.2800 metric tons are generated per year. SBE contains high percentage (%) of total solids ca.98, volatiles solids 35, oil content 30 and C: N ratio 258. Macronutrients expressed in percentage (%) were MgO 4.6, CaO 4.2, K2O 1.0, P2O 55.2, whereas as micronutrients were (%) F2O3 8.0 and MnO2 0.1. Mixing of cow dung manure with SBE at the rate of 50% on volatile solids bases improved methane yield by 9-fold (508ml/gVS compared to 55ml/gVs for only manure). The methane yield for only SBE was 16-fold (867 mL/gVS) compared to only manure (55ml/gVs). The SBE is therefore a cheap, abundantly available, excellent substrate for biogas production.

Open Access Original Research Article

Isolation and Identification of Endophyte Microorganisms from Bauhinia monandra Leaves, Mainly Actinobacteria

Rosilma de O. Araujo-Melo, Igor F. A. C. Souza, Carlos Vinícius J. de Oliveira, Janete M. de Araújo, Kêsia X. F. R. de Sena, Luana Cassandra Breitenbach Barroso Coel

Biotechnology Journal International, Page 1-12
DOI: 10.9734/BJI/2017/31636

The present work aimed the prospection of microorganisms from Bauhinia monandra leaves, with the purpose to identify endophytics to obtain strains with possible biotechnological applications. B. monandra leaves, disinfected with hypochlorite solution, were macerated in phosphate buffered saline and seeded in ten culture media containing antibacterial or antifungal agents. The endophytic filamentous fungus strains detected belonged to the genera Penicillium, Curvullaria and Aspergillus. Non-filamentous endophyte bacteria were grouped in the genera Bacillus, Burkholderia, Enterobacter and strains of endophytic Actinobacteria were classified as Streptomyces and Nocardiopsis. The isolation of endophytic microorganisms with nine culture media revealed better bacteria development with L-arginine agar; inorganic salt starch agar and potato dextrose agar were superior to Actinobacteria and fungus strains, respectively. The present study revealed the predominance of the genus Penicillium in leaves of B. monandra. This work introduces the first data of identification from endophyte Actinobacteria in the leaves of B. monandra.

Open Access Original Research Article

Heat Shock Proteins in Cassia Species

Geetika Pant, Ugam Kumari Chauhan, Sudhakar Malla, S. Banupriya, Rajesh Pati

Biotechnology Journal International, Page 1-10
DOI: 10.9734/BJI/2017/31006

Aim: Abiotic stresses, such as drought, salinity, extreme temperatures, chemical toxicity and oxidative stress, are serious threats to plants. Heat shock proteins can assist in protein refolding under stress conditions at molecular levels. The proposed study was aimed to analyze and characterize the heat shock proteins at the molecular level in the Cassia species.

Study Design: The experiments designed were intended to study and characterize the heat shock proteins in the Cassia species using molecular approach.

Methodology: The samples were subjected to the varying heat treatments (30⁰C, 37⁰C and 42⁰C), and the proteins obtained were further sequenced for similarity search. The peptide sequence was synthesized chemically and conjugated with Keyhole limpet hemocyanin (KLH) for the production of antibodies. Western blot was carried out with the polyclonal antibodies to confirm the results.

Results: The gel analysis revealed a clearly visible over-expressed band at the highest induced temperature (42ºC) depicting the presence of a heat shock response in Cassia. BLASTp search of the peptide sequences obtained from trypsin digestion followed by LC-MS resulted in 14 hits, out of which one of the peptide was similar to a known HSP, thioredoxin peroxidase from Nicotiana tabacum. Rabbit polyclonal antibodies were synthesized against the KLH-tagged peptide. The Western blot confirmed binding of probes to the heat shock band observed for the temperature treatment at 42ºC.

Conclusions: The obtained stress proteins can be used as molecular biomarkers for identifying the structural adaptive changes in plants based on their ecological habitats. The study can also help us in synthesizing the HSPs which could aid in protecting the plants against heat stress and maintain the homeostasis.