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We recently described the production of a detergent-biocompatible crude protease from Streptomyces mutabilis strain TN-X30. Here, we describe the purification, characterization, and immobilization of the serine alkaline protease (named SPSM), as well as the cloning, sequencing, and over-expression of its corresponding gene (spSM). Pure enzyme was obtained after ammonium sulphate precipitation followed by heat-treatment and Sephacryl® S-200 column purification. The sequence of the first 26 NH

Background Oleuropein, the main bitter phenolic glucoside responsible for green olive bitterness, may be degraded by the β-glucosidase enzyme to release glucose and phenolic compounds. Results Lactobacillus plantarum FSO1 and Candida pelliculosa L18 strains, isolated from natural fermented green olives, were tested for their β-glucosidase production and activity at different initial pH, NaCl concentrations, and temperature. The results showed that strains produced extracellular and induced β-glucosidase, with a molecular weight of 60 kD. The strains demonstrated their biodegradation capacity of oleuropein, associated with the accumulation of hydroxytyrosol and other phenolic compounds, resulting in antioxidant activity values significantly higher than that of ascorbic acid. The highest production value of β-glucosidase was 0.91 U/ml obtained at pH 5 and pH 6, respectively for L. plantarum FSO1 and C. pelliculosa L18. The increase of NaCl concentration, from 0 to 10% (w/v), inhibited the production of β-glucosidase for both strains. However, the β-glucosidase was activated with an increase of NaCl concentration, with a maximum activity obtained at 8% NaCl (w/v). The enzyme activity was optimal at pH 5 for both strains, while the optimum temperature was 45 °C for L. plantarum FSO1 and 35 °C for C. pelliculosa L18. Conclusions L. plantarum FSO1 and C. pelliculosa L18 strains showed their ability to produce an extracellular and induced β-glucosidase enzyme with promising traits for application in the biological processing of table olives.

Lactic acid bacteria (LAB) are ubiquitous bacteria associated with spontaneous lactic fermentation of vegetables, dairy and meat products. They are generally recognized as safe (GRAS), and they are involved in transformation of probiotic lacto-fermented foods, highly desired for their nutraceutical properties. The antifungal activity is one of the exciting properties of LAB, because of its possible application in food bio-preservation, as alternative to chemical preservatives. Many recent research works have been developed on antifungal activity of LAB, and they demonstrate their capacity to produce various antifungal compounds, (i.e. organic acids, PLA, proteinaceous compounds, peptides, cyclic dipeptides, fatty acids, and other compounds), of different properties (hydrophilic, hydrophobic and amphiphilic). The effectiveness of LAB in controlling spoilage and pathogenic fungi, demonstrated in different agricultural and food products, can be due to the synergistic effect between their antifungal compounds of different properties; where the amphiphilic-compounds allow the contact between the target microbial cell (hydrophilic compartment) and antifungal hydrophobic-compounds. Further studies on the interaction between compounds of these three properties are to de be developed, in order to highlight more their mechanism of action, and make LAB more profitable in improving shelf life and nutraceutical properties of foods.

The objective of this work is the study of the antifungal and antibacterial activity of Lactiplantibacillus plantarum S61 strains, isolated from traditional fermenting green olives against Rhodotorula glutinis UMP 22 and Listeria monocytogenes ATCC 19117, and its application in meat as bio-preservative agent. The cell-free supernatant (CFS) of Lpb. plantarum S61 shows high inhibition zones, which are 22.45 ± 0.49 and 17.75 ± 0.35 mm, against Rhodotorula glutinis and Listeria monocytogenes. The minimum fungicidal and bactericidal concentrations of the CFS obtained are 8% (v/v) and 10% (v/v), respectively. The competition assay, realized in liquid medium by co-culture of Lpb. plantarum S61 with Rho Rhodotorula glutinis and L. monocytogenes, led to inhibition percentages of 77.72% and 89.52%, respectively. However, the antimicrobial activity of Lpb. plantarum S61 was revealed a proteinaceous nature. Lpb. plantarum S61 strain allowed the reduction of L. monocytogenes in minced poultry meat during 7 days of storage at 4 °C. In addition, Lpb. plantarum S61 improved the physicochemical and color parameters of poultry minced meat. Lpb. plantarum S61 and/or its antimicrobial compounds can be applied as bio-preservative agent in meat product and food industry.

In Morocco, citrus fruits processing produces large quantities of waste and by-products, which represent 50% to 60% of the fresh mass of fruit. In eastern Morocco, generally citrus processing by-products ends up being thrown in the public landfill. These by-products are rich in bioactive compounds such as antioxidants and essential oils. The objective of this study is the chemical and biological characterization of essential oils extracted from citrus peels of four species, including Lemon (Citrus limon), Bitter orange (Citrus aurantium), Berkane clementine (Citrus Clementina) and Navel, a species of orange (Citrus sinensis). The efficiency of two extraction techniques used, hydro-distillation (Traditional method) and microwave assisted extraction (Eco-friendly green method), is highlighted by comparing yields of essential oils (EOs). Thus microwave assisted extraction green method has shown the highest yields ranging from 10,47% (Clementine) to 11,5% (Navel). While by hydrodistillation, the yield of (OE) did not exceed (5,95%). Besides, chemical composition and antioxidant activity of EOs of the four citrus were studied. Thus, the CPG-MS analyzes identified some twenty minor compounds and the D-limonene as the major constituent with contents generally greater than 55% and which reached almost (97,12%). In the case of EO of Lemon (Citrus lemon), the antioxidant activity was determined by the DPPH free-radical scavenging ability, the highest antioxidant activity (44,45%) is registered in the Navel’s EO, and the lowest was observed in the Bigarade’s EO (10,66%). In addition, antimicrobial activity was tested, EOs of lemon have shown the strongest antifungal power with values of 35 mm of antagonistic effect, but no antibacterial activity was detected for the four types of Eos. These EOs would be of interest in both agriculture and the food industry by acting as bio-fungicide but also as additives for compounds of medical and cosmetic interest.

Biological preservatives are increasingly in demand as an alternative to chemicals, because of consumer’s health and environmental concerns. Lactic acid bacteria isolated from fermented olives, considered as extreme environment, may encompass significant antimicrobial activity against microorganisms involved in foodborne diseases and food spoilage. Three LAB strains (Levilactobacillus brevis S27, Lactiplantibacillus pentosus S42, and Lactiplantibacillus plantarum S62), isolated from fermented olive, were studied for their antimicrobial activity against pathogenic bacteria (Listeria innocua and E. coli) and yeast (Rhodotorula glutinis), involved in foodborne diseases and food spoilage. The microbial culture, cell-free supernatant (CFS) of all LAB strains showed inhibition zone (IZ) values ranges of 26.33-28.83 mm, 14.16-16.33 mm and 10-12.16 mm against R. glutinis, L. innocua and E. coli, respectively. The IZ values ranges obtained with NCFS (neutralized CFS at pH 7) against R. glutinis and L. innocua were respectively of 17.83-20.50 mm and 13.16-14.50 mm, whereas no inhibition was obtained against E. coli. Their bioactive molecules of proteinaceous nature showed that among the four peptide bands obtained, two bands of approximately 42 and 56 kDa are common for the three LAB strains (S27, S42 and S62). Two additional bands of 46 and 89kD are observed in S42 and S27, respectively. The minimum inhibitory concentration of 10-fold concentrated CFS (10xCFS) obtained was 5% against E. coli and 4.5% against L. innocua and R. glutinis. The 10xCFS of Lb. plantarum S62, selected as the most efficient strain against pathogens, demonstrated its effectiveness in decontamination of milk at concentrations of 0.45% (v/v) and 0.5% (v/v) against L. innocua and E. coli. The CFS allowed, at 0.5 mL/piece of 5 cm diameter, a total inhibition of R. glutinis, without affecting the color of Pastilla leaf. The CFS of Lb. plantarum S62 can be used as a bio-preservative against pathogenic bacteria (L. innocua and E. coli) in milk, and against R. glutinis in Pastilla leaf.

The aim of the present study was to evaluate some technological properties and potential probiotic of 14 Lactobacillus strains isolated from brines of natural fermenting Moroccan Picholine green olive. The brine samples, collected from industrial environments, were analyzed for their Physico-chemical and microbiological properties. The Lactobacillus strains were characterized for their technological and physiological properties. The results obtained showed that the olive brines have an average pH of 4.32, an acidity of 0.61% and chloride contents of 6.42%. LAB and yeasts are the most dominant microorganisms in olive brine samples. The selected Lactobacillus (14 strains) showed low resistance to pH 2 and high resistance to bile salts (up to 2%), with values ranges from 1.98%-4.70% and 63.12%-86.48%, respectively. All the Lactobacillus strains displayed high levels of acidification and can produce β-glucosidase, protease, and cellulase in large amounts. The diacetyl production is detected in five Lactobacillus strains.