The Use of Essential Oil Embedded in Polylactic Acid/Chitosan-Based Film for Mango Post-Harvest Application against Pathogenic Fungi.

Mango has a high global demand. Fruit fungal disease causes post-harvest mango and fruit losses. Conventional chemical fungicides and plastic prevent fungal diseases but they are hazardous to humans and the environment. Direct application of essential oil for post-harvest fruit control is not a cost-effective approach. The current work offers an eco-friendly alternative to controlling the post-harvest disease of fruit using a film amalgamated with oil derived from Melaleuca alternifolia. Further, this research also aimed to assess the mechanical, antioxidant, and antifungal properties of the film infused with essential oil. ASTM D882 was performed to determine the tensile strength of the film. The antioxidant reaction of the film was assessed using the DPPH assay. In vitro and in vivo tests were used to evaluate the inhibitory development of the film against pathogenic fungi, by comparing the film with different levels of essential oil together with the treatment of the control and chemical fungicide. Disk diffusion was used to evaluate mycelial growth inhibition, where the film incorporated with 1.2 wt% essential oil yielded the best results. For in vivo testing of wounded mango, the disease incidence was successfully reduced. For in vivo testing of unwounded mango to which the film incorporated with essential oil was applied, although some quality parameters such as the color index were not significantly affected, weight loss was reduced, soluble solid content was increased, and firmness was increased, compared to the control. Thus, the film incorporated with essential oil (EO) from M. alternifolia can be an environmentally friendly alternative to the conventional approach and the direct application of essential oil to control post-harvest disease in mango.

Microwave-assisted solvent-free extraction of essential oil from Coleus aromaticus: anti-phytopathogenic potential for fruit post-harvesting.

This work evaluates the fungicidal effect of essential oil from Coleus aromaticus (C. aromaticus) by solvent-free microwave-assisted extraction with a yield of 0.54%. Fourier-transform infrared spectroscopy was utilised to identify the functional groups, which were O-H, C-O, C-H, and C=C. Gas chromatography-mass spectrometry analysis was performed to determine the primary essential oil components, namely, thymol (92.62%), thymoquinone (2.64%), creosol (1.77%), linalool (1.68%), p-Cymene-2,5-diol (0.73%), and p-Cymene (0.56%). The inhibitory effect of essential oil extracted from C. aromaticus against the isolated fungi, Aspergillus niger from mango, was investigated. The mycelial growth inhibition of the extracted essential oil by the poisoned food test and disc diffusion assay showed the reduction at 79.63 ± 1.7 and 70.45 ± 6.54%, respectively. In vivo experiment was conducted with artificially wounded and unwounded mangoes, applying the extracted essential oil to the wounded mangoes inoculated with A. niger that could decrease the disease incidence from 100 to 58.33 ± 14.43%. Meanwhile, the treatment of the extracted essential oil did not affect the quality of the mango and it also shows improvement in weight loss reduction of the mango in comparison to the chemical fungicide and untreated mango. Hence, essential oil from C. aromaticus by solvent-free microwave-assisted extraction could be used as an effective control for the fruit spoilage and potential source of fruit preservative.

Biosynthesis of butyl esters from crude oil of palm fruit and kernel using halophilic lipase secretion by Marinobacter litoralis SW-45.

Initially, a new moderate halophilic strain was locally isolated from seawater. The partial 16S rRNA sequence analysis positioned the organism in Marinobacter genus and was named 'Marinobacter litoralis SW-45'. This study further demonstrates successful utilization of the halophilic M. litoralis SW-45 lipase (MLL) for butyl ester synthesis from crude palm fruit oil (CPO) and kernel oil (CPKO) in heptane and solvent-free system, respectively, using hydroesterification. Hydrolysis and esterification of enzymatic [Thermomyces lanuginosus lipase (TLL)] hydrolysis of CPO and CPKO to free fatty acids (FFA) followed by MLL-catalytic esterification of the concentrated FFAs with butanol (acyl acceptor) to synthesize butyl esters were performed. A one-factor-at-a-time technique (OFAT) was used to study the influence of physicochemical factors on the esterification reaction. Under optimal esterification conditions of 40 and 45 °C, 150 and 230 rpm, 50% (v/v) biocatalyst concentration, 1:1 and 5:1 butanol:FFA, 9% and 15% (w/v) NaCl, 60 and 15 min reaction time for CPO- and CPKO-derived FFA esterification system, maximum ester conversion of 62.2% and 69.1%, respectively, was attained. Gas chromatography (GC) analysis confirmed the products formed as butyl esters. These results showed halophilic lipase has promising potential to be used for biosynthesis of butyl esters in oleochemical industry.

Whole genome sequence of moderate halophilic marine bacterium Marinobacter litoralis SW-45: Abundance of non-coding RNAs.

A report on the de novo Whole Genome Sequence (WGS) of Marinobacter litoralis SW-45, a moderately salt-tolerant bacterium isolated from the seawater in Malaysia is presented. The strain has a genome size of 3.45 Mb and is capable of producing halophilic lipase, protease and esterase enzymes. Computational prediction of non-coding RNA (ncRNA) genes in M. litoralis SW-45 was performed using standalone software known as the non-coding RNA characterization (nocoRNAc). In addition, a phylogenetic tree showing the evolutionary relationship between the strain and other members of the genus Marinobacter was constructed using 16SrRNA sequence information. A total of 385 ncRNA transcripts, 1124 terminator region, and 2350 Stress Induced Duplex Destabilization sites were predicted. The current WGS shotgun project has provided the relevant genetic information that may be useful for the strain's improvement studies. This manuscript gives the first description of M. litoralis with a complete genome.

Isolation and Partial Characterisation of Thermophilic Cellulolytic Bacteria from North Malaysian Tropical Mangrove Soil.

This study reports the biodiversity of thermophilic cellulolytic bacterial strains that present in the north Malaysian mangrove ecosystem. Soil samples were collected at the four most northern state of Malaysia (Perak, Pulau Pinang, Kedah and Perlis). The samples obtained were first enriched in nutrient broth at 45°C and 55°C prior culturing in the carboxymethylcellulose (CMC) agar medium. Repeated streaking was performed on the CMC agar to obtain a pure culture of each isolate prior subjecting it to hydrolysis capacity testing. The isolates that showing the cellulolytic zone (halozone) were sent for 16S rRNA sequencing. Total seven isolates (two from Perak, three from Kedah, another two were from Perlis and Penang each) showed halozone. The isolate (KFX-40) from Kedah exhibited highest halozone of 3.42 ± 0.58, meanwhile, the one obtained from Perak (AFZ-0) showed the lowest hydrolysis capacity (2.61 ± 0.10). Based on 16S rRNA sequencing results, 5 isolates (AFY-40, AFZ-0, KFX-40, RFY-20, and PFX-40) were determined to be Anoxybacillus sp. The other two isolates were identified as Bacillus subtilis (KFY-40) and Paenibacillus dendritiformis (KFX-0). Based on growth curve, doubling time of Anoxybacillus sp. UniMAP-KB06 was calculated to be 32.3 min. Optimal cellulose hydrolysis temperature and pH of this strain were determined to be 55°C and 6.0 respectively. Addition of Mg2+ and Ca2+ were found to enhance the cellulase activity while Fe3+ acted as an enzyme inhibitor.

Improvement of halophilic cellulase production from locally isolated fungal strain.

Halophilic cellulases from the newly isolated fungus, Aspergillus terreus UniMAP AA-6 were found to be useful for in situ saccharification of ionic liquids treated lignocelluloses. Efforts have been taken to improve the enzyme production through statistical optimization approach namely Plackett-Burman design and the Face Centered Central Composite Design (FCCCD). Plackett-Burman experimental design was used to screen the medium components and process conditions. It was found that carboxymethylcellulose (CMC), FeSO4·7H2O, NaCl, MgSO4·7H2O, peptone, agitation speed and inoculum size significantly influence the production of halophilic cellulase. On the other hand, KH2PO4, KOH, yeast extract and temperature had a negative effect on enzyme production. Further optimization through FCCCD revealed that the optimization approach improved halophilic cellulase production from 0.029 U/ml to 0.0625 U/ml, which was approximately 2.2-times greater than before optimization.

Applicability evaluation of Deep Eutectic Solvents-Cellulase system for lignocellulose hydrolysis.

Deep Eutectic Solvents (DESs) have recently emerged as a new generation of ionic liquids for lignocellulose pretreatment. However, DESs contain salt components which tend to inactivate cellulase in the subsequent saccharification process. To alleviate this problem, it is necessary to evaluate the applicability of the DESs-Cellulase system. This was accomplished in the present study by first studying the stability of cellulase in the presence of selected DESs followed by applicability evaluation based on glucose production, energy consumption and kinetic performance. Results showed that the cellulase was able to retain more than 90% of its original activity in the presence of 10% (v/v) for glycerol based DES (GLY) and ethylene glycol based DES (EG). Furthermore, both DESs system exhibited higher glucose percentage enhancement and lower energy consumption as compared to diluted alkali system. Among the two DESs studied, EG showed comparatively better kinetic performance.

Potential halophilic cellulases for in situ enzymatic saccharification of ionic liquids pretreated lignocelluloses.

Ionic liquids (ILs) have been used as an alternative green solvent for lignocelluloses pretreatment. However, being a salt, ILs exhibit an inhibitory effect on cellulases activity, thus making the subsequent saccharification inefficient. The aim of the present study is to produce salt-tolerant cellulases, with the rationale that the enzyme also tolerant to the presence of ILs. The enzyme was produced from a locally isolated halophilic strain and was characterized and assessed for its tolerance to different types of ionic liquids. The results showed that halophilic cellulases produced from Aspergillus terreus UniMAP AA-6 exhibited higher tolerance to ILs and enhanced thermo stability in the presence of high saline conditions.

Effect of steaming process on new formulation and physical properties of earthworm-based fish pellets for African catfish (Clarias gariepinus).

Fish feed has been recognized as one of main part/unit in aquaculture industry. However, current fish feed faces few challenges in terms of health aspects and cost issues. Alternatively, new nutritional and economical/low cost formulation of fish pellets was designed by combination of earthworm powder and other economical ingredients such as fishmeal, soybean waste, rice bran and tapioca flour. The formulation was calculated using Pearson's square and optimized by One-Factor-At-Time (OFAT) method. The effect of steaming processing on the water stability, soaking experiment, protein leaching test and breaking force of the earthworm-based fish pellets was investigated. Results indicate steam pellet at 80 degrees C for 40 min has higher water stability, less protein leaching and more durable than unsteam pellets. Introduction of this new formulation of fish meal is expected to provide essential nutrient, energy and improved the quality of pellets to fuel the growth of aquaculture industry.

Optimization of complex fermentation media for glucose oxidase production using statistical approach.

Production cost of enzyme is largely determined by the type of the strain and raw material used to propagate the strain. Hence, selection of the strain and raw materials is crucial in enzyme production. For Glucose oxidase (GOx), previous studies showed Aspergillus terreus UniMAP AA-1 offers a better alternative to the existing sources. Thus, a lower production cost could be logically anticipated by growing the strain in a cheaper complex media such as molasses. In this work, sugar cane molasses, supplemented with urea and carbonate salt and a locally isolated strain Aspergillus terreus UniMAP AA-1 were used to produce a crude GOx enzyme in a small scale. A statistical optimization approach namely Response Surface Methodology (RSM) was used to optimize the media components for highest GOx activity. It was found that the highest GOx activity was achieved using a combination of molasses, carbonate salt and urea at concentration 32.51, 4.58 and 0.93% (w/v), respectively. This study provides an alternative optimized media conditions for GOx production using locally available raw materials.