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Fulltext Isolation and Characterization of Lactobacillus spp. from Kefir Samples in Malaysia

Talib N, Mohamad NE, Yeap SK, Hussin Y, Aziz MNM, Masarudin MJ, et al.

Molecules, 2019 Jul 17;24(14).
PMID: 31319614 DOI: 10.3390/molecules24142606

Kefir is a homemade, natural fermented product comprised of a probiotic bacteria and yeast complex. Kefir consumption has been associated with many advantageous properties to general health, including as an antioxidative, anti-obesity, anti-inflammatory, anti-microbial, and anti-tumor moiety. This beverage is commonly found and consumed by people in the United States of America, China, France, Brazil, and Japan. Recently, the consumption of kefir has been popularized in other countries including Malaysia. The microflora in kefir from different countries differs due to variations in culture conditions and the starter media. Thus, this study was aimed at isolating and characterizing the lactic acid bacteria that are predominant in Malaysian kefir grains via macroscopic examination and 16S ribosomal RNA gene sequencing. The results revealed that the Malaysian kefir grains are dominated by three different strains of Lactobacillus strains, which are Lactobacillus harbinensis, Lactobacillusparacasei, and Lactobacillus plantarum. The probiotic properties of these strains, such as acid and bile salt tolerances, adherence ability to the intestinal mucosa, antibiotic resistance, and hemolytic test, were subsequently conducted and extensively studied. The isolated Lactobacillus spp. from kefir H maintained its survival rate within 3 h of incubation at pH 3 and pH 4 at 98.0 ± 3.3% and 96.1 ± 1.7% of bacteria growth and exhibited the highest survival at bile salt condition at 0.3% and 0.5%. The same isolate also showed high adherence ability to intestinal cells at 96.3 ± 0.01%, has antibiotic resistance towards ampicillin, penicillin, and tetracycline, and showed no hemolytic activity. In addition, the results of antioxidant activity tests demonstrated that isolated Lactobacillus spp. from kefir G possessed high antioxidant activities for total phenolic content (TPC), total flavonoid content (TFC), ferric reducing ability of plasma (FRAP), and 1,1-diphenyl-2-picryl-hydrazine (DPPH) assay compared to other isolates. From these data, all Lactobacillus spp. isolated from Malaysian kefir serve as promising candidates for probiotics foods and beverage since they exhibit potential probiotic properties and antioxidant activities.

Matched MeSH terms: Lactobacillus/growth & development; Lactobacillus/isolation & purification*; Lactobacillus/chemistry; Lactobacillus plantarum/growth & development; Lactobacillus plantarum/isolation & purification*; Lactobacillus plantarum/chemistry
Fulltext Characterization, yield optimization, scale up and biopreservative potential of fermencin SA715, a novel bacteriocin from Lactobacillus fermentum GA715 of goat milk origin

Wayah SB, Philip K

Microb Cell Fact, 2018 Aug 13;17(1):125.
PMID: 30103750 DOI: 10.1186/s12934-018-0972-1

BACKGROUND: Emergence of antibiotic resistance and growing consumer trend towards foods containing biopreservatives stimulated the search for alternative antimicrobials. This research is aimed at characterizing, investigating the mechanism of action, scale up optimization and evaluating the biopreservative potential of a bacteriocin from Lactobacillus fermentum.
RESULTS: Fermencin SA715 is a novel, broad-spectrum, non-pore-forming and cell wall-associated bacteriocin isolated from L. fermentum GA715 of goat milk origin. A combination of hydrophobic interaction chromatography, solid-phase extraction and reversed-phase HPLC was necessary for purification of the bacteriocin to homogeneity. It has a molecular weight of 1792.537 Da as revealed by MALDI-TOF mass spectrometry. Fermencin SA715 is potent at micromolar concentration, possesses high thermal and pH stability and inactivated by proteolytic enzymes thereby revealing its proteinaceous nature. Biomass accumulation and production of fermencin SA715 was optimum in a newly synthesized growth medium. Fermencin SA715 did not occur in the absence of manganese(II) sulphate. Tween 80, ascorbic acid, sodium citrate and magnesium sulphate enhanced the production of fermencin SA715. Sucrose is the preferred carbon source for growth and bacteriocin production. Sodium chloride concentration higher than 1% suppressed growth and production of fermencin SA715. Optimum bacteriocin production occurred at 37 °C and pH 6-7. Scale up of fermencin SA715 production involved batch fermentation in a bioreactor at a constant pH of 6.5 which resulted in enhanced production. Fermencin SA715 doubled the shelf life and improved the microbiological safety of fresh banana. Bacteriocin application followed by refrigeration tripled the shell life of banana.
CONCLUSIONS: This study reveals the huge potential of fermencin SA715 as a future biopreservative for bananas and reveals other interesting characteristics which can be exploited in the preservation of other foods. Furthermore insights on the factors influencing the production of fermencin SA715 have been revealed and optimized condition for its production has been established facilitating future commercial production.

Matched MeSH terms: Lactobacillus fermentum/metabolism*
Fulltext Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins

Jalilsood T, Baradaran A, Song AA, Foo HL, Mustafa S, Saad WZ, et al.

Microb Cell Fact, 2015;14:96.
PMID: 26150120 DOI: 10.1186/s12934-015-0283-8

Bacterial biofilms are a preferred mode of growth for many types of microorganisms in their natural environments. The ability of pathogens to integrate within a biofilm is pivotal to their survival. The possibility of biofilm formation in Lactobacillus communities is also important in various industrial and medical settings. Lactobacilli can eliminate the colonization of different pathogenic microorganisms. Alternatively, new opportunities are now arising with the rapidly expanding potential of lactic acid bacteria biofilms as bio-control agents against food-borne pathogens.

Matched MeSH terms: Lactobacillus/physiology
Screening of lactobacillus strains against salmonella both isolated from malaysian free-range chicken intestine for use as probiotic

Wan Mohtar Wan Yusoff, Hutari A, Jaseem WS, Aidil Abdul Hamid

Sains Malaysiana, 2011;40:1115-1122.

A total of eight strains of Lactobacillus and two strains of Salmonella were isolated from free-range Malaysian chickens intestine. Evaluation based on in vitro studies included aggregation, co-aggregation, growth with bile salts, tolerance to acidic pH, and inhibitory activity were carried out. The isolated Lactobacillus were Lactobacillus fermentum IA, Lactobacillus fermentum IB, Lactobacillus fermentum IC, Lactobacillus fermentum ID, Lactobacillus salivarius subsp. salicinus IE, Lactobacillus salivarius subsp. salicinus IF, Lactobacillus salivarius subsp. salivarius IG, and Lactobacillus spp. IH. The corresponding isolated Salmonella were Salmonella spp. 3B21 and Salmonella spp. 1A12. The ability of aggregation and also tolerance to pH 2.5 are found in Lactobacillus fermentum ID, Lactobacillus salivarius subsp. salicinus IF, Lactobacillus salivarius subsp. salivarius IG, and Lactobacillus spp. IH. The isolate most resistance to 1% bile salts is Lactobacillus fermentum ID but observed to be weak in inhibitory activity against Salmonella spp. The best co-aggregation and strongest inhibitory activity against Salmonella spp. was observed in Lactobacillus salivarius subsp. salivarius IG. Despite being not so resistant in the presence of bile salts 0.5 and 1% (w/v), the lag time in the presence of bile salts 0.3% (w/v) of Lactobacillus salivarius subsp. salivarius IG and also for Lactobacillus spp. IH are the shortest. Based on good aggregation properties, the best co-aggregation, tolerance to acidic pH 2.5 and bile salts 0.3% (w/v) and strongest inhibitory activity against Salmonella spp., Lactobacillus salivarius subsp. salivarius IG comes out as the best candidate as probiotic for chicken.

Matched MeSH terms: Lactobacillus; Lactobacillus fermentum; Lactobacillus salivarius
Ultrasound enhanced growth and cholesterol removal of Lactobacillus fermentum FTDC 1311 in the parent cells but not the subsequent passages

Lye HS, Khoo BY, Karim AA, Rusul G, Liong MT

Ultrason Sonochem, 2012 Jul;19(4):901-8.
PMID: 22265020 DOI: 10.1016/j.ultsonch.2011.12.018

The aim of this study was to evaluate the effect of ultrasound on the intestinal adherence ability, cell growth, and cholesterol removal ability of parent cells and subsequent passages of Lactobacillus fermentum FTDC 1311. Ultrasound significantly decreased the intestinal adherence ability of treated parent cells compared to that of the control by 11.32% (P<0.05), which may be due to the protein denaturation upon local heating. Growth of treated parent cells also decreased by 4.45% (P<0.05) immediately upon ultrasound (0-4h) and showed an increase (P<0.05) in the viability by 2.18-2.34% during the later stage of fermentation (12-20 h) compared to that of the control. In addition, an increase (P<0.05) in assimilation of cholesterol (>9.74%) was also observed for treated parent cells compared to that of the control, accompanied by increased (P<0.05) incorporation of cholesterol into the cellular membrane. This was supported by the increased ratio of membrane cholesterol:phospholipids (C:P), saturation of cholesterol in the apolar regions, upper phospholipids regions, and polar regions of membrane phospholipids of parent cells compared to that of the control (P<0.05). However, such traits were not inherited by the subsequent passages of treated cells (first, second, and third passages). Our data suggested that ultrasound treatment could be used to improve cholesterol removal ability of parent cells without inducing permanent damage/defects on treated cells of subsequent passages.

Matched MeSH terms: Lactobacillus fermentum/cytology; Lactobacillus fermentum/growth & development*; Lactobacillus fermentum/chemistry*
Isolation and characterization of a novel Lactobacillus plantarum MMB-07 from traditional Suanyu for Acanthogobius hasta fermentation

Yang J, Lu J, Zhu Q, Tao Y, Zhu Q, Guo C, et al.

J Biosci Bioeng, 2021 Aug;132(2):161-166.
PMID: 33972168 DOI: 10.1016/j.jbiosc.2020.12.016

As one of Lianyungang's most famous specialties, Acanthogobius hasta is delicious and nutritious fish, but is extremely susceptible to spoilage during transportation and storage. In this study, Lactobacillus plantarum MMB-07 was isolated from traditional fermented sour fish to reduce losses and improve the utilization and food value of A. hasta. L. plantarum MMB-07 had good ability of acid production and acid resistance. Moreover, it could also inhibit common pathogens in food or aquatic products to ensure the safety of fermented products. MMB-07 was used to ferment A. hasta and obtain fermented Suanyu rich in nutrition value and good flavor. The volatile base nitrogen was 18.44 mg/100 g and the fermented fish meat maintained second-grade freshness. Thiobarbituric acid assay was 0.90 mg/kg and fat in fish meat was oxidized to a low degree. The studies indicated that MMB-07 has a high application prospect in low salt fermented fish.

Matched MeSH terms: Lactobacillus plantarum*
Fulltext Some physico-chemical qualities and acceptability of fermented curry paste

Abolhassani, Y, Khan, M.A., Salam, A.B, Ghasem, M.

International Food Research Journal, 2009;16(4):581-588.
MyJurnal

The effects of lactic acid bacteria (Lactobacillus plantarum and Lactobacillus bulgaricus) inoculation
on the sensory attributes and consumers acceptance of fermented curry paste compared with uncultured sample were assessed. pH, titratable acidity (TA) and color changes, during four-month storage were monitored. Hedonic test was utilized to evaluate consumer perception and acceptability of fermented and ordinary curry pastes. Rapid pH drop was observed in inoculated sample with Lb. plantarum presenting better performance than the Lb. bulgaricus. Titratable acidity increased significantly (p0.05) in most of the attributes of
original recipe and fermented curry paste except for color and sweetness. In summary, this study showed fermented curry paste with Lb. plantarum and Lb. bulgaricus exhibited new sensory attributes encouraging acceptability by consumers.

Matched MeSH terms: Lactobacillus; Lactobacillus plantarum
Fulltext Antifungal Prospect of Bacillus cereus Postbiotic on Crustacean Pathogen, Lagenidium thermophilum

Elliecpearl Jasca J, Annita Seok KY, Suraini L, Chun YA, Julian R, Sano M, et al.

Biocontrol Sci, 2021;26(4):201-205.
PMID: 35013016 DOI: 10.4265/bio.26.201

Pathogenic marine fungi, Lagenidium thermophilum is known causative agent in the crustacean industry. Current disinfection practice in hatchery has risks and negative impacts which prompts suitable substitute to synthetic antifungal agents. Thus, this study was conducted to evaluate the antifungal potential of postbiotic from four potential probiotics towards marine oomycetes, L. thermophilum IPMB 1401. The screening test showed that the Lactobacillus plantarum GS12 and Bacillus cereus GS15 postbiotics were positive for antifungal activity on L. thermophilum IPMB 1401. These two bacterial extracts have minimum inhibitory concentration （MIC） at 50%. The toxicity assay on MIC level of the postbiotic revealed that the cumulative mortality of brine shrimp nauplii exposed to B. cereus postbiotic was significantly lower compared to L. plantarum GS12 postbiotic and formalin. This indicates a high potential of B. cereus GS15 as a prospect for alternative control method for fungal infections in the crustacean culture industry.

Matched MeSH terms: Lactobacillus plantarum*
Analysis of CRISPR-Cas Loci and their Targets in Levilactobacillus brevis

Goh YX, Wang M, Hou XP, He Y, Ou HY

Interdiscip Sci, 2023 Sep;15(3):349-359.
PMID: 36849628 DOI: 10.1007/s12539-023-00555-1

The CRISPR‒Cas system acts as a bacterial defense mechanism by conferring adaptive immunity and limiting genetic reshuffling. However, under adverse environmental hazards, bacteria can employ their CRISPR‒Cas system to exchange genes that are vital for adaptation and survival. Levilactobacillus brevis is a lactic acid bacterium with great potential for commercial purposes because it can be genetically manipulated to enhance its functionality and nutritional value. Nevertheless, the CRISPR‒Cas system might interfere with the genetic modification process. Additionally, little is known about the CRISPR‒Cas system in this industrially important microorganism. Here, we investigate the prevalence, diversity, and targets of CRISPR‒Cas systems in the genus Levilactobacillus, further focusing on complete genomes of L. brevis. Using the CRISPRCasFinder webserver, we identified 801 putative CRISPR-Cas systems in the genus Levilactobacillus. Further investigation focusing on the complete genomes of L. brevis revealed 54 putative CRISPR-Cas systems. Of these, 46 were orphan CRISPRs, and eight were CRISPR‒Cas systems. The type II-A CRISPR‒Cas system is the most common in Levilactobacillus and L. brevis complete genomes. Analysis of the spacer's target showed that the CRISPR‒Cas systems of L. brevis mainly target the enterococcal plasmids. Comparative analysis of putative CRISPR-Cas loci in Levilactobacillus brevis.

Matched MeSH terms: Lactobacillus brevis*
Enhancing food preservation with postbiotic metabolites γ-aminobutyric acid (GABA) and bacteriocin-like inhibitory substances (BLIS) produced by Lactobacillus brevis C23 co-cultures in plant-based medium

Chuah WW, Tan JS, Hazwani Oslan SN, Bothi Raja P

Prep Biochem Biotechnol, 2024 Apr;54(4):514-525.
PMID: 37694843 DOI: 10.1080/10826068.2023.2252047

Lactic acid bacteria (LAB) can produce γ-aminobutyric acid (GABA) with antioxidant properties and sedative effects when it binds to the GABA receptor in the human brain. LAB can also produce bacteriocin-like inhibitory substances (BLIS) with antimicrobial capabilities during carbohydrate fermentation. GABA and BLIS are natural compounds with potential health benefits and food preservation properties. Lactobacillus brevis C23 was co-cultured with three different LABs as inducers, which produced the highest GABA content and BLIS activity. They were cultured in various plant-based media to obtain an edible and better-tasting final product over commercially available media like MRS broth. A coconut-based medium with additives was optimized using response surface methodology (RSM) to increase GABA and BLIS production. The optimized medium for maximum GABA production (3.22 ± 0.01 mg/mL) and BLIS activity (84.40 ± 0.44%) was a 5.5% coconut medium containing 0.23% glucose, 1.44% Tween 20, 0.48% L-glutamic acid, and 0.02% pyridoxine. Due to the presence of GABA, the cell-free supernatant (CFS) as a postbiotic showed higher antioxidant activity than other food preservatives like nisin and potassium sorbate. Finally, microbiological tests on food samples showed that the postbiotic was more effective than other preservatives at combating the growth of LAB, molds and coliform bacteria, making it a possible food preservative.

Matched MeSH terms: Lactobacillus brevis*
Electroporation enhances the ability of lactobacilli to remove cholesterol

Lye HS, Karim AA, Rusul G, Liong MT

J Dairy Sci, 2011 Oct;94(10):4820-30.
PMID: 21943733 DOI: 10.3168/jds.2011-4426

The objective of the present study was to evaluate the effect of electroporation on the membrane properties of lactobacilli and their ability to remove cholesterol in vitro. The growth of lactobacilli cells treated at 7.5 kV/cm for 4 ms was increased by 0.89 to 1.96 log(10) cfu/mL upon fermentation at 37 °C for 20 h, the increase being attributed to the reversible and transient formation of pores and defragmentation of clumped cells. In addition, an increase of cholesterol assimilation as high as 127.2% was observed for most cells electroporated at a field strength of 7.5 kV/cm for 3.5 ms compared with a lower field strength of 2.5 kV/cm. Electroporation also increased the incorporation of cholesterol into the cellular membrane, as shown by an increased cholesterol:phospholipids ratio (50.0-59.6%) upon treatment at 7.5 kV/cm compared with treatment at 2.5 kV/cm. Saturation of cholesterol was observed in different regions of the membrane bilayer such as upper phospholipids, apolar tail, and polar heads, as indicated by fluorescence anisotropy using 3 fluorescent probes. Electroporation could be a useful technique to increase the ability of lactobacilli to remove cholesterol for possible use as cholesterol-lowering adjuncts in the future.

Matched MeSH terms: Lactobacillus/growth & development; Lactobacillus/metabolism; Lactobacillus/physiology*; Lactobacillus/ultrastructure
Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk upon ultrasound-treatment

Ewe JA, Wan Abdullah WN, Bhat R, Karim AA, Liong MT

Ultrason Sonochem, 2012 Jan;19(1):160-73.
PMID: 21775184 DOI: 10.1016/j.ultsonch.2011.06.013

This study aimed at utilizing ultrasound treatment to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli (Lactobacillus acidophilus BT 1088, L. fermentum BT 8219, L. acidophilus FTDC 8633, L. gasseri FTDC 8131) were treated with ultrasound (30 kHz, 100 W) at different amplitudes (20%, 60% and 100%) for 60, 120 and 180 s prior to inoculation and fermentation in biotin-soymilk. The treatment affected the fatty acids chain of the cellular membrane lipid bilayer, as shown by an increased lipid peroxidation (P<0.05). This led to increased membrane fluidity and subsequently, membrane permeability (P<0.05). The permeabilized cellular membranes had facilitated nutrient internalization and subsequent growth enhancement (P<0.05). Higher amplitudes and longer durations of the treatment promoted growth of lactobacilli in soymilk, with viable counts exceeding 9 log CFU/mL. The intracellular and extracellular β-glucosidase specific activities of lactobacilli were also enhanced (P<0.05) upon ultrasound treatment, leading to increased bioconversion of isoflavones in soymilk, particularly genistin and malonyl genistin to genistein. Results from this study show that ultrasound treatment on lactobacilli cells promotes (P<0.05) the β-glucosidase activity of cells for the benefit of enhanced (P<0.05) isoflavone glucosides bioconversion to bioactive aglycones in soymilk.

Matched MeSH terms: Lactobacillus/growth & development*; Lactobacillus/chemistry
Improved production of live cells of Lactobacillus rhamnosus by continuous cultivation using glucose-yeast extract medium

Ling LS, Mohamad R, Rahim RA, Wan HY, Ariff AB

J Microbiol, 2006 Aug;44(4):439-46.
PMID: 16953180

In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates (0.05 h(-1) to 0.40 h(-1)) using a 2 L stirred tank fermenter with a working volume of 600 ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, mu-max, was estimated at 0.40 h(-1), and the Monod cell growth saturation constant, Ks, at approximately 0.25 g/L. Maximum cell viability (1.3 x 10(10) CFU/ml) was achieved in the dilution rate range of D = 0.28 h(-1) to 0.35 h(-1). Both maximum viable cell yield and productivity were achieved at D = 0.35 h(-1). The continuous cultivation of L. rhamnosus at D = 0.35 h(-1) resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.

Matched MeSH terms: Lactobacillus rhamnosus/growth & development*; Lactobacillus rhamnosus/metabolism
Adhesion ability and cytotoxic evaluation of lactobacillus strains isolated from malaysian fermented fish (pekasam) on ht-29 and ccd-18co intestinal cells

Ida Muryany, Ahmad Rohi Ghazali, Nor Fadilah Rajab, Hing HL, Ina-Salwany, Mohd Zamri Saad, et al.

Sains Malaysiana, 2018;47:2391-2399.

Bacterial adhesion to host cells is the most important probiotic character. However, the adhesion of probiotic should not
affect the viability of the host cells. In this study, Lactobacillus plantarum strain L8, Lactobacillus plantarum strain L20
and Lactobacillus pentosus strain S1 were tested for their cytotoxic effects through MTT assay and their ability to adhere
and colonize on HT-29 and CCD-18Co intestinal cells as detected microscopically using light microscopy and Scanning
Electron Microscopy (SEM). No cytotoxicity effects were observed on both intestinal cells following 24 h treatment with
all Lactobacillus strains. Additionally, all strains demonstrated strong adhesive activity where more than 100 bacteria
adhered to both intestinal cells although differences in the adhesion scores observed among different strains. The adhesion
as observed via SEM showed an autoagreggative pattern and adhered as clusters on the surface of both intestinal cells.
In conclusion, all three Lactobacillus strains are non-cytotoxic to both cells with strong adhesion ability on intestinal
cells and this study also proved that Malaysian fermented fish are good source of probiotic bacteria.

Matched MeSH terms: Lactobacillus; Lactobacillus plantarum; Lactobacillus pentosus
Microencapsulation of Lactobacillus rhamnosus GG with flaxseed mucilage using co-extrusion technique

Lai K, How Y, Pui L

J Microencapsul, 2021 Mar;38(2):134-148.
PMID: 33306440 DOI: 10.1080/02652048.2020.1863490

AIM: This study aimed to evaluate the protective effect of flaxseed mucilage on the co-extrusion microencapsulation of Lactobacillus rhamnosus GG.
METHODS: Core flow rate, chitosan coating, and flaxseed mucilage concentration were optimised for the microencapsulation of L. rhamnosus. The microbeads were characterised and evaluated on microencapsulation efficiency and cell released after 6 h of sequential digestion.
RESULTS: The optimised parameters for the L. rhamnosus microencapsulation were 1.0 mL/min core flow rate, 0.4% (w/v) chitosan coating, and 0.4% (w/v) flaxseed mucilage. The L. rhamnosus microbeads with flaxseed mucilage in core and wall materials had a smooth surface with 781.3 µm diameter, the highest microencapsulation efficiency (98.8% w/w), lowest swelling (5196.7% w/w) and erosion ratio (515.5% w/w), and least cell release (<40% w/w) with 9.31 log10 CFU mL-1 after sequential digestion.
CONCLUSIONS: This study showed the protective capacity of flaxseed mucilage towards the L. rhamnosus GG during microencapsulation and gastrointestinal environment.

Matched MeSH terms: Lactobacillus rhamnosus/cytology*; Lactobacillus rhamnosus/metabolism
Purification of exopolysaccharide produced from Lactobacillus spp. using ionic-liquid as adjuvant in alcohol/salt-based aqueous two-phase system for its antidiabetic properties

Raj ST, Puspanadan S, Gan CY, Tan JS

Int J Biol Macromol, 2024 May;267(Pt 2):131376.
PMID: 38608981 DOI: 10.1016/j.ijbiomac.2024.131376

Diabetes is a chronic, metabolic disease characterized by hyperglycemia resulting from either insufficient insulin production or impaired cellular response to insulin. Exopolysaccharides (EPS) produced by Lactobacillus spp. demonstrated promising therapeutic potential in terms of their anti-diabetic properties. Extraction and purification of EPS produced by Lactobacillus acidophilus and Limosilactobacillus reuteri were performed using ethanol precipitation, followed by alcohol/salt based aqueous two-phase system (ATPS). The purification process involved ethanol precipitation followed by an alcohol/salt-based ATPS. The study systematically investigated various purification parameters in ATPS, including ethanol concentration, type and concentration of ionic liquid, type and concentration of salt and pH of salt. Purified EPS contents from L. acidophilus (63.30 μg/mL) and L. reuteri (146.48 μg/mL) were obtained under optimum conditions of ATPS which consisted of 30 % (w/w) ethanol, 25 % (w/w) dipotassium hydrogen phosphate at pH 10 and 2 % (w/w) 1-butyl-3-methylimidazolium octyl sulfate. The extracted EPS content was determined using phenol sulphuric acid method. In α-amylase inhibition tests, the inhibitory rate was found to be 92.52 % (L. reuteri) and 90.64 % (L. acidophilus), while in α-glucosidase inhibition tests, the inhibitory rate was 73.58 % (L. reuteri) and 68.77 % (L. acidophilus), based on the optimized parameters selected in ATPS. These results suggest that the purified EPS derived from the postbiotics of Lactobacillus spp. hold promise as potential antidiabetic agents.

Matched MeSH terms: Lactobacillus*; Lactobacillus acidophilus
Effect of temperature and pH on the probiotication of Punica granatum juice using Lactobacillus species

Mustafa SM, Chua LS, El-Enshasy HA, Abd Majid FA, Hanapi SZ, Abdul Malik R

J Food Biochem, 2019 04;43(4):e12805.
PMID: 31353583 DOI: 10.1111/jfbc.12805

This study was focused on the effects of fermentation temperature and pH on the quality of Punica granatum juice probioticated with Lactobacillus species: Lactobacillus plantarum, Lactobacillus casei, Lactobacillus bulgaricus, and Lactobacillus salivarius. The whole fruit juice of P. granatum which is rich with phytonutrients appeared to be a good probiotic carrier. The probiotication was carried out for 24 hr at 30, 35, and 37°C and pH 2.5, 4.0, and 5.5 under microaerophilic conditions. The results found that P. granatum juice cultivated with L. casei had a better growth profile with a higher biomass density at 37°C around pH 3.5-4.0. Probiotication could maintain the scavenging activity of P. granatum juice cultivated with L. casei. The scavenging activity achieved up to 90% inhibition at the concentration of 5 mg/ml. The whole fruit-squeezed P. granatum juice was suitable for the growth of Lactobacillus species even without supplementation during cultivation. PRACTICAL APPLICATIONS: The findings of this study presented the potential of P. granatum juice (whole fruit) to be used as a good probiotic carrier, particularly for Lactobacillus species without supplementation. High nutritious P. granatum juice catered the need of probiotic bacteria during fermentation. Probiotication could maintain the antioxidant capacity of the juice in term of its radical scavenging activity. The antioxidant capacity was mainly attributed to the metabolites such as phenolic acids (romarinic acid and caftaric acid) and flavonoids (quercetin, quercetin 3-glucoside, rutin and kaempferol rutinoside). With the optimized temperature (37°C) and pH (4.00), probiotic bacteria could growth well up to a cell viability of 2.46 × 1010 cfu/ml. This offers P. granatum to be developed into functional food to cater to the needs of the consumers who are lactose intolerant to dairy products.

Matched MeSH terms: Lactobacillus casei/growth & development; Lactobacillus casei/metabolism*; Lactobacillus plantarum/growth & development; Lactobacillus plantarum/metabolism*
Physio-chemical, microbiological properties of tempoyak and molecular characterisation of lactic acid bacteria isolated from tempoyak

Chuah LO, Shamila-Syuhada AK, Liong MT, Rosma A, Thong KL, Rusul G

Food Microbiol, 2016 Sep;58:95-104.
PMID: 27217364 DOI: 10.1016/j.fm.2016.04.002

This study aims to determine physio-chemical properties of tempoyak, characterise the various indigenous species of lactic acid bacteria (LAB) present at different stages of fermentation and also to determine the survival of selected foodborne pathogens in tempoyak. The predominant microorganisms present in tempoyak were LAB (8.88-10.42 log CFU/g). Fructobacillus durionis and Lactobacillus plantarum were the dominant members of LAB. Other LAB species detected for the first time in tempoyak were a fructophilic strain of Lactobacillus fructivorans, Leuconostoc dextranicum, Lactobacillus collinoides and Lactobacillus paracasei. Heterofermentative Leuconostoc mesenteroides and F. durionis were predominant in the initial stage of fermentation, and as fermentation proceeded, F. durionis remained predominant, but towards the end of fermentation, homofermentative Lb. plantarum became the predominant species. Lactic, acetic and propionic acids were present in concentrations ranging from 0.30 to 9.65, 0.51 to 7.14 and 3.90 to 7.31 mg/g, respectively. Genotyping showed a high degree of diversity among F. durionis and Lb. plantarum isolates, suggesting different sources of LAB. All tested Lb. plantarum and F. durionis (except for one isolate) isolates were multidrug resistant. Salmonella spp., Listeria monocytogenes and Staphylococcus aureus were not detected. However, survival study showed that these pathogens could survive up to 8-12 days. The results aiming at improving the quality and safety of tempoyak.

Matched MeSH terms: Lactobacillus/genetics; Lactobacillus/isolation & purification*; Lactobacillus plantarum/genetics; Lactobacillus plantarum/isolation & purification
Characteristics of Metroxylon sagu resistant starch type III as prebiotic substance

Zi-Ni T, Rosma A, Napisah H, Karim AA, Liong MT

J Food Sci, 2015 Apr;80(4):H875-82.
PMID: 25739421 DOI: 10.1111/1750-3841.12817

Resistant starch type III (RS3 ) was produced from sago (Metroxylon sagu) and evaluated for its characteristics as a prebiotic. Two RS3 samples designated sago RS and HCl-sago RS contained 35.71% and 68.30% RS, respectively, were subjected to hydrolyses by gastric juice and digestive enzymes and to absorption. Both sago RS and HCl-sago RS were resistant to 180 min hydrolysis by gastric acidity at pH 1 to 4 with less than 0.85% hydrolyzed. Both samples were also resistant toward hydrolysis by gastrointestinal tract enzymes and intestinal absorption with 96.75% and 98.69% of RS3 were recovered respectively after 3.5 h digestion and overnight dialysis at 37 °C. Sago RS3 supported the growth of both beneficial (lactobacilli and Bifidobacteria) and pathogenic microbes (Escherichia coli, Campylobacter coli, and Clostridium perfringens) in the range of 2.60 to 3.91 log10 CFU/mL. Hence, prebiotic activity score was applied to describe the extent to which sago RS3 supports selective growth of the lactobacilli and bifidobacteria strains over pathogenic bacteria. The highest scores were obtained from Bifidobacterium sp. FTDC8943 grown on sago RS (+0.26) and HCl-sago RS (+0.24) followed by L. bulgaricus FTDC1511 grown on sago RS (+0.21). The findings had suggested that sago RS3 has the prebiotic partial characteristics and it is suggested to further assess the suitability of sago RS3 as a prebiotic material.

Matched MeSH terms: Lactobacillus/growth & development*
Fulltext Multiplex Real-Time PCR with HRM for Detection of Lactobacillus sakei and Lactobacillus curvatus in Food Samples

Kurbakov KA, Konorov EA, Minaev MY, Kuznetsova OA

Food Technol Biotechnol, 2019 Mar;57(1):97-104.
PMID: 31316281 DOI: 10.17113/ftb.57.01.19.5983

Optimization of fermentation processes requires monitoring the species composition of starter cultures and their growth during fermentation. Most starter cultures contain closely related species. Nowadays, high-resolution melting (HRM) analysis is extensively used for multiplex identification of closely related species. In the present paper, we applied real-time polymerase chain reaction (PCR) with HRM analysis for the detection and differentiation of Lactobacillus sakei and L. curvatus. A primer pair was selected for the site of the rpoA gene of Lactobacillus spp. Eleven starter cultures and fifteen fermented sausages with a known bacterial composition were successfully tested using real-time PCR with HRM analysis with the developed primer pair.

Matched MeSH terms: Lactobacillus; Lactobacillus sakei

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