Flavor Precursors and Volatile Compounds Improvement of Unfermented Cocoa Beans by Hydrolysis Using Bromelain.

Cocoa fermentation is an essential process that produces flavor precursors. However, many small farmers in Indonesia directly dry their cocoa beans without fermentation due to low yield and long fermentation time, resulting in fewer flavor precursors and cocoa flavor. Therefore, this study aimed to enhance the flavor precursors, particularly free amino acids and volatile compounds, of unfermented cocoa beans by hydrolysis, using bromelain. Unfermented cocoa beans were previously hydrolyzed with bromelain at concentrations of 3.5, 7, and 10.5 U/mL for 4, 6, and 8 h, respectively. An analysis of enzyme activity, degree of hydrolysis, free amino acids, reducing sugar, polyphenols, and volatile compounds was then conducted using unfermented and fermented cocoa beans as negative and positive controls, respectively. The results showed that the highest degree of hydrolysis was 42.95% at 10.5 U/mL for 6 h, although it was not significantly different from the hydrolysis at 3.5 U/mL for 8 h. This indicates a higher reducing sugar and lower polyphenols content than unfermented cocoa beans. There was also an increase in free amino acids, especially hydrophobic amino acids, such as phenylalanine, valine, leucine, alanine, and tyrosine, and desirable volatile compounds, such as pyrazines. Therefore, this suggests that hydrolysis with bromelain increased the flavor precursors and cocoa-bean flavors.

Correlation between the chemical, microbiological and sensory characteristics of cream cheese using a mixed and single probiotic culture.

The use of mixed culture in the fermentation industry requires more complicated equipment, processes, and monitoring systems; therefore, a single culture may be preferable. This study aimed to investigate the correlation between chemical and microbiological properties and sensory characteristics. In addition, this study aimed to determine the different characteristics of cheese made using single probiotic cultures of Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Kita-3 and mixed culture. The obtained results showed that the chemical characteristics of cream cheese made using single cultures were similar to those of cream cheese made using mixed cultures. The viability of the cells remained high after 35 days of storage for the single culture, which was not the case for the mixed culture. In terms of sensory analysis, cream cheese made using the single culture L. plantarum Kita-3 showed higher overall liking score among the samples, which might correlate with the high ester and ketone content. In addition, there was a strong relationship between the ethyl octanoate and methyl butanol contents and the overall liking score. The results of this study showed that the use of a single culture of L. plantarum Kita-3 could improve the sensory characteristics of cream cheese with probiotic properties. This study also contributed to the development of cream cheese production, particularly in the screening of potential starters.

Gut Microbiota Composition in Undernourished Children Associated with Diet and Sociodemographic Factors: A Case-Control Study in Indonesia.

Malnutrition, which consists of undernutrition and overnutrition, is associated with gut microbiota composition, diet, and sociodemographic factors. Undernutrition is a nutrient deficiency that that should be identified to prevent other diseases. In this study, we evaluate the gut microbiota composition in undernourished children in association with diet and sociodemographic factors. We observed normal children (n= 20) and undernourished children (n= 20) for ten days in Lombok and Yogyakarta. Diet, sociodemographic factors, and medical records were recorded using food records, screening forms, and standard household questionnaires. Gut microbiota analysis was performed using 16S rRNA gene sequencing targeting the V3-V4 region. The result showed that the undernourished group had lower energy intake. In addition, the undernourished group had lower quality of medical records, parent knowledge, education, and exclusive breastfeeding. Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, and Verrucomicrobia were significantly different between normal and undernourished children. Based on LefSe, we determined that Akkermansia is a biomarker for undernourished children. In conclusion, diet and sociodemographic factors affect the gut microbiota composition of undernourished children.

Enzymatic Glycerolysis of Palm Kernel Olein-Stearin Blend for Monolaurin Synthesis as an Emulsifier and Antibacterial.

Monolaurin is a monoacylglycerol, which can act as an emulsifier and antibacterial. Palm kernel oil is a monolaurin raw material that can be fractionated into palm kernel olein (PKOo) and palm kernel stearin (PKS). Therefore, this study prepares monolaurin through enzymatic glycerolysis of the PKOo-PKS blend. The effects of enzyme concentration, molar ratio of oil to glycerol, solvent to oil ratio, and reaction temperature on the products of glycerolysis were investigated. The best conditions were selected for further production, purification, and characterization of the monolaurin. The results showed that the best glycerolysis condition was obtained with an enzyme concentration of 10% w/w, an oil-glycerol molar ratio of 1:4, a solvent-oil ratio of 2:1 v/w, and a glycerolysis temperature of 40 °C with a stirring speed of 600 rpm based on the monoacylglycerol (MAG) concentration. The identification of the sample with FTIR and NMR indicated that the purified glycerolysis product is the monolaurin. The thermal analysis showed a large endothermic peak with a melting point of 35.56 °C. The purified monolaurin has a HLB value of 5.92, and an emulsion capacity and stability of 93.66 ± 1.85% and 89.54 ± 3.36%, respectively. The minimum inhibitory concentration (MIC) of the monolaurin for Escherichia coli FNCC 0091 and Staphylococcus aureus FNCC 0047 were at 500 ppm, and 100 ppm for Bacillus subtilis FNCC 0060.

Safety assessment of the indigenous probiotic strain Lactiplantibacillus plantarum subsp. plantarum Kita-3 using Sprague-Dawley rats as a model.

Lactiplantibacillus plantarum subsp. plantarum Kita-3 is a candidate probiotic from Halloumi cheese produced by Mazaraat Artisan Cheese, Yogyakarta, Indonesia. This study evaluated the safety of consuming a high dose of L. plantarum subsp. plantarum Kita-3 in Sprague-Dawley rats for 28 days. Eighteen male rats were randomly divided into three groups, such as the control group, the skim milk group, and the probiotic group. Feed intake and body weight were monitored, and blood samples, organs (kidneys, spleen, and liver), and the colon were dissected. Organ weight, hematological parameters, serum glutamic oxaloacetic transaminase (SGOT), and serum glutamic pyruvic transaminase (SGPT) concentrations, as well as intestinal morphology of the rats, were measured. Microbial analyses were carried out on the digesta, feces, blood, organs, and colon. The results showed that consumption of L. plantarum did not negatively affect general health, organ weight, hematological parameters, SGOT and SGPT activities, or intestinal morphology. The number of L. plantarum in the feces of rats increased significantly, indicating survival of the bacterium in the gastrointestinal tract. The bacteria in the blood, organs, and colon of all groups were identified using repetitive-polymerase chain reaction with the BOXA1R primers and further by 16S rRNA gene sequencing analysis, which revealed that they were not identical to L. plantarum subsp. plantarum Kita-3. Thus, this strain did not translocate to the blood or organs of rats. Therefore, L. plantarum subsp. plantarum Kita-3 is likely to be safe for human consumption.

Adhesion Properties of Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7 on Sprague Dawley Rat Intestine.

Adhesion capacity is considered one of the selection criteria for probiotic strains. The purpose of this study was to determine the adhesion properties of two candidate probiotics, Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7. The evaluation included the hydrophobicity of the cell surface using microbial adhesion to hydrocarbons (MATH), autoaggregation, and the adhesion of L. plantarum Dad-13 and L. plantarum Mut-7 to the intestinal mucosa of Sprague Dawley rat, followed by genomic analysis of the two L. plantarum strains. L. plantarum Dad-13 and L. plantarum Mut-7 showed a high surface hydrophobicity (78.9% and 83.5%) and medium autoaggregation ability (40.9% and 57.5%, respectively). The exposure of both isolates to the surface of the rat intestine increased the total number of lactic acid bacteria on the colon compartment, from 2.9 log CFU/cm2 to 4.4 log CFU/cm2 in L. plantarum Dad-13 treatment and to 3.86 log CFU/cm2 in L. plantarum Mut-7 treatment. The results indicate the ability of two L. plantarum to attach to the surface of the rat intestine. The number of indigenous E. coli in the colon also decreased when the compartment was exposed to L. plantarum Dad-13 and Mut-7, from 2.9 log CFU/cm2 to 1 log CFU/cm2. Genomic analysis revealed that both strains have genes related to adhesion properties that could play an important role in increasing the adherence of probiotics to the intestinal mucosa such as gene encoding fibronectin-binding protein, chaperonin heat shock protein 33 (Hsp33), and genes related to the capsule and cell wall biosynthesis. Based on these findings, we believe that L. plantarum Dad-13 and L. plantarum Mut-7 have adhesion properties to the intestinal mucosa in the rat intestine model system. The present research will be essential to elucidate the molecular mechanism associated with adhesion in our two probiotic strains.

Effect of probiotic Lactobacillus plantarum Dad-13 powder consumption on the gut microbiota and intestinal health of overweight adults.

BACKGROUND: Shifting on lifestyle, diet, and physical activity contributed on increasing number of obese people around the world. Multiple factors influence the development of obesity. Some research suggested that gut microbiota (GM) plays an important role in nutrient absorption and energy regulation of individuals, thus affecting their nutritional status. Report of Indonesia Basic Health Research showed that the prevalence of obesity in every province tended to increase. Although the root cause of obesity is excessive calorie intake compared with expenditure, the differences in gut microbial ecology between healthy and obese humans may affect energy homeostasis. GM affect body weight, especially obesity. Probiotics that are consumed while alive and able to colonize in the intestine are expected to increase the population of good bacteria, especially Bifidobacteria and Lactobacilli, and suppress pathogens such as Enterobacteriaceae and Staphylococcus. The strain of L. plantarum Dad-13 has been demonstrated to survive and colonize in the gastrointestinal tract of healthy Indonesian adults who consume fermented milk containing L. plantarum Dad-13. The consumption of probiotic L. plantarum Dad-13 powder decreased E. coli and non-E. coli coliform bacteria in school-aged children in Indonesia. L. plantarum is a dominant bacterium in the average Indonesian's GM. For this reason, this bacterium is probably a more suitable probiotic for Indonesians. AIM: To determine the effect of the consumption of indigenous probiotic Lactobacillus plantarum Dad-13 powder in overweight adults in Yogyakarta (Indonesia). METHODS: Sixty overweight volunteers with a body mass index (BMI) equal to or greater than 25 consume indigenous probiotic powder L. plantarum Dad-13 (2 × 109 CFU/gram/sachet) for 90 d. The study was a randomized, double-blind, placebo-controlled study. The volunteers filled in a diary on a daily basis, which consisted of questions on study product intake (only during ingestion period), other food intake, number of bowel movements, fecal quality (consistency and color), any medications received, and any symptom of discomfort, such as diarrhea, constipation, vomiting, gassing, sensation of illness, etc. Fecal samples and the subjects' diaries were collected on the morning of day 10 + 1, which was marked as the end of the baseline period and the start of the ingestion period. During the ingestion period (from day 11 to day 101), several parameters to measure and analyze the results included body weight and height (once a month), the lipid profile, GM analysis using MiSeq, short-chain fatty acid (SCFA) analysis using gas chromatography, and the measurement of fecal pH using a pH meter. RESULTS: The consumption of indigenous probiotic powder L. plantarum Dad-13 caused the average body weight and BMI of the probiotic group to decrease from 84.54 ± 17.64 kg to 83.14 ± 14.71 kg and 33.10 ± 6.15 kg/m2 to 32.57 ± 5.01 kg/m2, respectively. No significant reduction of body weight and BMI in the placebo group was observed. An analysis of the microbiota showed that the number of Bacteroidetes, specifically Prevotella, increased significantly, while that of Firmicutes significantly decreased. No significant change in lipid profile in both groups was found. Also, no significant change in SCFAs (e.g., butyrate, propionate, acetic acid) and pH level was found after the consumption of the probiotic. CONCLUSION: No significant differences in pH before and after ingestion were observed in both the probiotic and placebo groups as well as in the lipid profile of both cholesterol and triglyceride, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and the LDL/HDL ratio. In addition, no significant changes in the concentration of SCFAs (e.g., acetic acid, propionate, and butyrate) were found after con-sumption. Interestingly, a significant decrease in body weight and BMI (P < 0.05) was determined in the treatment group. An analysis of GM shows that L. plantarum Dad-13 caused the Firmicutes population to decrease and the Bacteroidetes population (especially Prevotella) to increase.

Enzymatic glycerolysis-interesterification of palm stearin-olein blend for synthesis structured lipid containing high mono- and diacylglycerol.

The objective of this research was to evaluate enzymatic glycerolysis-interesterification to synthesize structured lipids (SLs) containing high monoacylglycerol (MAG) and diacylglycerol (DAG) from a palm stearin-olein blend (PS-PO blend). The results showed that the optimum conditions for the solvent to fat ratio, glycerol to fat ratio, and enzyme concentration were 2:1 (v/w), 1.5:1, and 15% (w/w), respectively. The conversion rate of MAG and DAG decreased at a high glycerol to fat ratio, low solvent to fat ratio, and high enzyme concentration due to an increase in viscosity and low agitation effectiveness. The emulsion capacity and stability of the SLs were 60.19% and 96.80%, respectively. The hardness of the SLs increased about 3.1-fold. The MAG, DAG, and triacylglycerol conversion rates were 0.45, 0.48, and 1.02%/h, respectively. Thus, glycerolysis-interesterification of a PS-PO blend increased DAG and MAG concentrations and further improved the hardness, emulsion capacity, and emulsion stability of the SLs.

Gut microbiota profile in healthy Indonesians.

BACKGROUND: Recently, gut microbiota has been associated with various diseases other than intestinal disease. Thus, there has been rapid growth in the study of gut microbiota. Considering the numerous factors influencing gut microbiota such as age, diet, etc., area-based research is required. Indonesia has numerous different tribes and each of these tribes have different lifestyles. Hence, it is expected that each tribe has a specific gut microbiota. A deeper insight into the composition of gut microbiota can be used to determine the condition of gut microbiota in Indonesians and to consider which treatment may be suitable and effective to improve health status. AIM: To investigate the gut microbiota of Indonesian subjects represented by Javanese and Balinese tribes by analyzing fecal samples. METHODS: Fecal samples were collected from a total of 80 individuals with 20 in each of the young groups ranging from 25-45 years and the elderly group aged 70 years or more from two different regions, Yogyakarta and Bali. Fecal sample collection was performed at the end of the assessment period (day 14 ± 1 d) during which time the subjects were not allowed to consume probiotic or antibiotic products. The quantification of various Clostridium subgroups, Lactobacillus subgroups, Enterococcus, Streptococcus, Staphylococcus, Bacteroides fragilis group and Prevotella, Bifidobacterium and Atopobium cluster, Enterobacteriaceae and Pseudomonas was performed using the Yakult intestinal flora-scan (YIF-SCAN). RESULTS: The bacterial population in younger subjects' feces was higher than that in the elderly population, with a total of approximately 10.0 - 10.6 log10 bacterial cells/g feces. The most abundant bacteria in all groups were Clostridium, followed by Prevotella, Atopobium, Bifidobacterium and Bacteroides. In the elderly, an increase in Enterobacteriaceae, Coliform and Escherichia coli was found. In terms of bacterial counts in Yogyakarta, total bacteria, Clostridium coccoides (C. coccoides) group, Bifidobacterium, Prevotella, Lactobacillus plantarum subgroup, and Streptococcus were significantly higher (P < 0.05) in younger than elderly subjects, while the Lactobacillus gasseri subgroup, Lactobacillus casei subgroup, and Lactobacillus reuteri subgroup counts were significantly lower (P < 0.05) in younger subjects. In Balinese subjects, total bacteria, C. coccoides group, Clostridium leptum subgroup, Bacteroides fragilis group, and Prevotella were significantly higher (P < 0.05) in younger compared to elderly individuals, while the Lactobacillus ruminis subgroup, and Enterobacteriaceae were significantly lower (P < 0.05) in younger subjects. The results also revealed that, besides the C. coccoides group and Clostridium leptum group being the most abundant gut microbiota in both Yogyakarta and Balinese people, the latter was indicated by a higher Clostridium perfringens count, which was almost 10 times that of Yogyakarta subjects. This may be a response to different lifestyles in the different tribes; however, this phenomenon requires further extensive study. CONCLUSION: Bacterial populations were higher in younger than in elderly subjects. Most abundant bacterial groups were Clostridium, Prevotella, Atopobium, Bifidobacterium, and Bacteroides. The level of Clostridium perfringens in Yogyakarta subjects was lower than that in Balinese subjects.