Characterization and shelf life of synbiotic drink powder from porang (Amorphophallus muelleri).

Amorphophallus muelleri BI was included in the Araceae family, which is a type of tuber. It is a tuber with high potential due to its abundant bioactive compounds. Amorphophallus muelleri BI flour (AF) contains a high glucomannan and carbon compounds that serve as nutrients for probiotic bacteria. Although Amorphophallus muelleri BI thrives in Indonesia, its utilization rate in the country remains relatively low and haven't been any studies conducted regarding synbiotic powder from AF. The primary objective of this research is to develop a synergistic beverage enriched with varying concentrations of Amorphophallus muelleri BI as a prebiotic and LA as probiotic (synbiotic). The process starts with culture preparation, synbiotic drink process, synbiotic and microencapsulation, includes the examination of solubility, proximate analysis, calorie content, viability, and shelf life. Results showed that the proximate and solubility had no significant effect. Synbiotic drink powder from AF can be produced using spray dry technology. The highest LA growth was observed when augmenting the AF quantity at a 0.4% concentration, which can be seen from the viability parameter with a value of 7.29 log CFU/g. Samples shelf life at -21 and 3 °C with LA viability critical parameter was determined to be 4 days.

Dietary supplementation of black rice bran to colon carcinogen-induced mice: Examining the development of colorectal cancer by improving environmental colon conditions.

This research aims to identify the effects of the administration of a black rice bran diet on colorectal cancer in dextran sodium sulfate and azoxymethane-induced BALB/c mice. The research was conducted on three groups consisting of eight Balb/c mice: two groups were fed with carcinogens, and the third group, referred to as the normal group, was supplied with Isotonic NaCl 0.9% intraperitoneally. One group fed with carcinogens was supplied a standard AIN 1993 M diet modified with black rice bran as a substitute of fibre source, while the other two mice groups were fed the standard diet (AIN-93M) containing cellulose fibre. At the 17th week, all mice were euthanized; their colonic sections were taken for histopathological evaluation, and cecum for short-chain fatty acids concentration, total lactic acid bacteria, pH and ß-glucuronidase activity evaluations. The results show an increase in the total lactic acid bacteria and short-chain fatty acids in the mice group fed with rice bran. Consequently, pH value and ß-glucuronidase activity had decreased. Histopathological evaluation of mucosal tissue exhibited inhibition of the tumor growth rate in the mice groups fed rice bran compared to the group supplied with the standard diet. Furthermore, the proliferating cell nuclear antigen expression had decreased significantly, while expression of caspase-8 and caspase-3 had increased notably, in the group fed with a rice bran diet. These results suggest that black rice bran can effectively inhibit colon carcinogenesis. The potential of black rice bran as a source of fibre has not been studied in detail regarding the inhibition mechanism of colorectal cancer cells; further investigation in this field could provide valuable information about new strategies to prevent colorectal cancer. This strand of research is very important to developing preventive methods against cancer and promoting the concept of healthy products, including functional foods.

The Occurrence of Folate Biosynthesis Genes in Lactic Acid Bacteria from Different Sources.

Research background: Lactic acid bacteria (LAB) are known to produce folate. However, this ability is highly strain-dependent. Folate synthesis in specific LAB strains is affected by the availability of folate, which can be consumed by other LAB under certain conditions. Moreover, differences in folate synthesis capabilities are related to the presence of folate biosynthesis-related genes and regulation of this pathway. Experimental approach: As basic information to better understand the regulation of folate biosynthesis among different LAB species and strains, folate biosynthetic genes were screened and identified in folate-producing and non-folate-producing LAB isolated from various local food sources in Indonesia. The extracellular folate productivity amounts of the isolates were analyzed using high-performance liquid chromatography with a diode array detector (HPLC-DAD). Results and conclusions: Eleven of the thirteen tested LAB isolates had all of the eight genes involved in folate biosynthesis (folE, folQ, folB, folK, folP, folC1, folA and folC2). Furthermore, these isolates produced extracellular folate ranging from 10.37 to 31.10 µg/mL. In contrast, two non-folate-producing isolates lacked several folate biosynthetic genes, such as folQ, folP and folA, which is possibly the reason for their inability to synthesize folate de novo. Phylogenetic tree construction revealed that the folate biosynthetic genes (excluding folK and folP) from six distinct species of folate-producing LAB isolates were monophyletic with homologous genes from other LAB species in the database. Novelty and scientific contribution: In this study, the distribution of folate biosynthetic genes in various LAB species was determined. The findings from this research support the use of folate biosynthesis marker genes in the genotypic screening for folate-producing LAB.

Hypothetical Regulation of Folate Biosynthesis and Strategies for Folate Overproduction in Lactic Acid Bacteria.

Folate (vitamin B9) is an essential nutrient for cell metabolism, especially in pregnant women; however, folate deficiency is a major global health issue. To address this issue, folate-rich fermented foods have been used as alternative sources of natural folate. Lactic acid bacteria (LAB), which are commonly involved in food fermentation, can synthesize and excrete folate into the medium, thereby increasing folate levels. However, screening for folate-producing LAB strains is necessary because this ability is highly dependent on the bacterial strain. Some strains of LAB consume folate, and their presence in a fermentation mix can lower the folate levels of the final product. Since microorganisms efficiently regulate folate biosynthesis to meet their growth needs, some strains of folate-producing LAB can deplete folate levels if folate is available in the media. Such folate-efficient producers possess a feedback inhibition mechanism that downregulates folate biosynthesis. Therefore, the application of folate-overproducing strains may be a key strategy for increasing folate levels in media with or without available folate. Many studies have been conducted to screen folate-producing bacteria, but very few have focused on the identification of overproducers. This is probably because of the limited understanding of the regulation of folate biosynthesis in LAB. In this review, we discuss the roles of folate-biosynthetic genes and their contributions to the ability of LAB to synthesize and regulate folate. In addition, we present various hypotheses regarding the regulation of the feedback inhibition mechanism of folate-biosynthetic enzymes and discuss strategies for obtaining folate-overproducing LAB strains.

Angiotensin-I-Converting Enzyme Inhibitory Peptides in Goat Milk Fermented by Lactic Acid Bacteria Isolated from Fermented Food and Breast Milk.

In this study, angiotensin-I-converting enzyme inhibitory (ACEI) activity was evaluated in fermented goat milk fermented by lactic acid bacteria (LAB) from fermented foods and breast milk. Furthermore, the potential for ACEI peptides was identified in fermented goat milk with the highest ACEI activity. The proteolytic specificity of LAB was also evaluated. The 2% isolate was inoculated into reconstituted goat milk (11%, w/v), then incubated at 37°C until pH 4.6 was reached. The supernatant produced by centrifugation was analyzed for ACEI activity and total peptide. Viable cell counts of LAB and titratable acidity were also evaluated after fermentation. Peptide identification was carried out using nano liquid chromatography mass spectrometry (LC-MS/MS), and potential as an ACEI peptide was carried out based on a literature review. The result revealed that ACEI activity was produced in all samples (20.44%-60.33%). Fermented goat milk of Lc. lactis ssp. lactis BD17 produced the highest ACEI activity (60.33%; IC50 0.297±0.10 mg/mL) after 48 h incubation, viable cell counts >8 Log CFU/mL, and peptide content of 4.037±0.27/mL. A total of 261 peptides were released, predominantly derived from casein (93%). The proteolytic specificity of Lc. lactis ssp. lactis BD17 through cleavage on the amino acid tyrosine, leucine, glutamic acid, and proline. A total of 21 peptides were identified as ACEI peptides. This study showed that one of the isolates from fermented food, namely Lc. lactis ssp. lactis BD17, has the potential as a starter culture for the production of fermented goat milk which has functional properties as a source of antihypertensive peptides.

Folate in Milk Fermented by Lactic Acid Bacteria from Different Food Sources.

Folates are essential micronutrients, and folate deficiency still occurs in many countries. Lactic acid bacteria (LAB) are known to be able to synthesize folates during fermentation, but the folate production is strain-dependent and influenced by the fermentation medium, presence of a folate precursor, and fermentation time. This study aimed to screen extracellular folate-producing LAB from local food sources and evaluate the factors influencing their folate biosynthesis during milk fermentation. The selection of folate-producing LAB was based on their ability to grow in folate-free medium (FACM), with folate concentrations quantified by microbiological assay. Growth of the 18 LAB in FACM varied between isolates, with only 8 isolates growing well and able to synthesize extracellular folate at relatively high concentrations (up to 24.27 ng/mL). The isolates with highest extracellular folate levels, Lactobacillus fermentum JK13 from kefir granules, Lactobacillus plantarum 4C261 from salted mustard, and Lactobacillus rhamnosus R23 from breast milk, were applied to milk fermentation. The last two isolates were probiotic candidates. The three isolates consumed folate when it was present in the milk, and its consumption was in line with their growth. The availability of folate precursors affected the amount of folate consumed, but did not lead to increased folate concentrations in the medium after 72 h fermentation. The results of this study indicate that these isolates cannot be utilized for producing folate in folate-containing milk, as it shows feedback inhibition on folate biosynthesis.

Lactobacillus rhamnosus Reduces Blood Glucose Level through Downregulation of Gluconeogenesis Gene Expression in Streptozotocin-Induced Diabetic Rats.

Some lactic acid bacteria (LAB) are observed to be potential probiotics with functional properties such as lowering fasting blood glucose (FBG), as a promising hyperglycemia management. This study investigated the ability and mechanism of Lactobacillus rhamnosus BSL and Lactobacillus rhamnosus R23 on lowering FBG in diabetic rats induced by streptozotocin (STZ). The rats were orally administered with L. rhamnosus BSL and L. rhamnosus R23 by giving 1 mL cell suspension (109 CFU/mL) daily for 30 days. The body weight (BW) was recorded once in three days, and FBG was recorded once in six days. An oral glucose tolerance test (OGTT) was measured 1 week after injection with STZ and before sacrifice. Fecal samples were collected on days 0, 15, and 30 for LAB population and identification, performed by PCR detecting 16S rRNA. Oral administration of L. rhamnosus BSL and L. rhamnosus R23 decreased FBG and improved glucose tolerance via downregulation of glucose-6-phosphatase (G6pc) expression by 0.57- and 0.60-fold change, respectively (P < 0.05). The lipid profiles, BUN, creatinine, SGOT, and SGPT were significantly (P < 0.05) different between normal and diabetic rats, but they were not significantly (P > 0.05) different among diabetic rats. Both strains were effective in increasing fecal LAB population. Molecular identification of the isolated LAB from fecal sample indicated that they were able to survive and pass through the digestive tract. These results suggested that both strains have the ability to manage blood glucose level and become a promising agent to manage hyperglycemia and diabetes.

Angiotensin-I-converting enzyme inhibitory peptides in milk fermented by indigenous lactic acid bacteria.

BACKGROUND AND AIM: Fermented milk can be used to produce antihypertensive peptides. Lactic acid bacteria (LAB) with its proteolytic system hydrolyze milk protein during fermentation to produce several peptides, which include antihypertensive bioactive peptides. This study aimed to investigate the ability of indigenous LAB for the production of angiotensin-I-converting enzyme inhibitory (ACE-I) peptides in fermented milk and to characterize the ACEI peptides. MATERIALS AND METHODS: Reconstituted milk (11%) inoculated with ten LAB isolates, and then incubated at 37°C until it reaches pH 4.6. The evaluation was carried out for LAB count, lactic acid concentration, peptide content, and ACE-I activity. The low molecular weight (MW) peptides (<3 kDa) were identified using Nano LC Ultimate 3000 series system Tandem Q Exactive Plus Orbitrap high-resolution mass spectrometry. RESULTS: The result showed that the ten LAB isolates were able to produce ACE-I in fermented milk with the activities in the range of 22.78±2.55-57.36±5.40%. The activity of ACE-I above 50% produced by Lactobacillus delbrueckii BD7, Lactococcus lactis ssp. lactis BD17, and Lactobacillus kefiri YK4 and JK17, with the highest activity of ACE-I produced by L. kefiri YK4 (IC50 0.261 mg/mL) and L. kefiri JK17 (IC50 0.308 mg/mL). Results of peptide identification showed that L. kefiri YK 4 could release as many as 1329, while L. kefiri JK 17 could release 174 peptides. The peptides produced were 95% derived from casein. The other peptides were from ά-lactalbumin, ß-lactoglobulin, and serum amyloid A. The peptides produced consisted of 6-19 amino acid residues, with MWs of 634-2079 Dalton and detected at 317-1093 m/z. A total of 30 peptides have been recognized based on literature searches as ACE-I peptides (sequence similarity: 100%). CONCLUSION: L. kefiri YK4 and JK17 are the potential to be used as starter cultures to produce the bioactive peptide as ACE-I in fermented milk.

In Vitro Characterization of Lactic Acid Bacteria from Indonesian Kefir Grains as Probiotics with Cholesterol-Lowering Effect.

Indonesian kefir grains are potential sources of lactic acid bacteria (LAB) that may act as probiotics with specific functional properties. In this study we explored the potential of the probiotic and cholesterol-lowering effect of LAB isolated from Indonesian kefir grains obtained from Bogor, Bandung, Jakarta, and Yogyakarta. The results revealed that 10 isolates showed considerable survivability at low pH and bile salt with total cell reduction of ~3 log colony-forming units per milliliter after exposure to pH 2.5 and 0.5% (w/v) bile salt for 1 and 3 h, respectively. All strains exhibited strong antimicrobial activities against pathogenic bacteria and were sensitive to a wide spectrum of antibiotics but exhibited weak bile salt hydrolase activity. Identification based on 16S RNA suggested that nine isolates were Lactobacillus kefiri and one was Lactobacillus rhamnosus. The ability of the isolates to reduce cholesterol from the media varied, ranging from 22.08% to 68.75% with the highest reduction shown by L. kefiri JK17. The ability to remove cholesterol from the media decreased greatly in resting and dead cells, ranging from 14.58% to 22.08% in resting cells and from 7.89% to 18.17% in dead cells. It can be concluded that Indonesian kefir grains contain LAB potentially acting as probiotics capable of reducing cholesterol. The cholesterol-lowering effect especially occurs when the cells are metabolically active.

Nutritional status of lactating women in Bogor district, Indonesia: cross-sectional dietary intake in three economic quintiles and comparison with pre-pregnant women.

The way in which women accommodate for their increased nutritional needs during the lactation period is poorly investigated. In a cross-sectional study involving 220 lactating women (LW), equally distributed in economic quintiles (Q2, Q3, Q4), we investigated whether habitual dietary intake of LW differed from that of 200 pre-pregnant women (PPW) studied using the same methodology. Differences in dietary intake and nutrition sufficiency according to economic status were also investigated. Dietary intake data were collected using 2×24-h dietary recalls and FFQ. Energy, protein, Fe, Ca, Zn and vitamins A and C intakes were calculated utilising local food composition tables and were compared against Indonesian recommendations for adequacy. Energy and protein intakes 25 % of LW. For Q2-LW, this proportion was >50 %, except for protein. LW across all studied economic quintiles approximately doubled their vegetable intake, and 71 % of LW indicated a belief that this enhances lactation performance. Biochemical status parameters were analysed in a subset of forty-five women. Anaemia as well as Fe, Zn and Se deficiencies were prevalent among LW, supporting the nutrient intake deficiency data. Despite increasing intakes in LW compared with PPW, habitual diets in the study area do not provide for daily nutrient requirements in substantial proportions for both LW and PPW across all investigated economic groups.

Nutritional status of pre-pregnant and pregnant women residing in Bogor district, Indonesia: a cross-sectional dietary and nutrient intake study.

The diet of Indonesian women of childbearing age is relatively poor, posing increased risk for suboptimal pregnancy outcome. In a cross-sectional study including 403 women in three economic quintiles (Q), we investigated differences in dietary intake and nutrition sufficiency according to economic status and whether regular dietary intakes of pregnant women (PW, n 203) differ from that of pre-pregnant women (PPW, n 200). Dietary intake data were collected using 2×24 h dietary recall and FFQ. Energy, protein, Fe, Ca, Zn, vitamin A and vitamin C intakes were calculated utilising Food Composition Tables and compared with Indonesian recommendations for adequacy. Energy and protein intakes <70 % and Fe, Ca, Zn, vitamin A and vitamin C intakes <77 % of the local recommendation were considered insufficient. A higher intake of milk/dairy products (Q3, Q4), fruits and vegetables (Q2) and snacks (Q3) in PW was observed compared with PPW, but insufficient to meet nutrient requirements. Nutrient intake calculations showed insufficiency in 26 % of PPW for protein and up to 70 % for Fe. Deficient nutrient intakes in PW were found in 49 % of the subjects for energy and up to 85 % for Fe. Energy and protein intakes decreased with decreasing economic quintiles in PPW and PW. Ca and vitamin A intakes were lower in Q2 compared with Q4. The proportion of animal protein in Q2-PW was low (31 %). Biochemical status parameters in a subset showed that anaemia and Fe and Zn deficiencies were prevalent among PPW and PW. Habitual diets in the study area did not meet the nutrient requirements for both PPW and PW across the investigated economic groups.