Study of Oenological Fermentation: Which Strategy and Which Tools?

Wine fermentation is a specific and complex research subject and its control is essential to ensure full process completion while improving wine quality. It displays several specificities, in particular, (i) musts with a very high sugar content, low pH, and some limiting nutrients, as well as a great variability in must composition according to the year, grape variety, and so on; (ii) atypical fermentation conditions with non-isothermal temperature profiles, a quasi-anaerobiosis and legal constraints with a limited and predefined list of authorized operations. New challenges have emerged, related to the increasing diversity of commercially available yeast strains; the fluctuating composition of musts, particularly owing to climate change; and sustainability, which has become a key issue. This paper synthesizes approaches implemented to address all these issues. It details the example of our laboratory that, for many years, has been developing an integrated approach to study yeast diversity, understand their metabolism, and develop new fermentation control strategies. This approach requires the development of specific fermentation devices to study yeast metabolism in a controlled environment that mimics practical conditions and to develop original fermentation control strategies. All these tools are described here, together with their role in the overall scientific strategy and complementary approaches in the literature.

Production of Nigerian Yoghurt Using Lactic Acid Bacteria as Starter Cultures

Aim: This work was carried out to investigate the influence of Lactic Acid Bacteria (LAB) on organoleptic quality and proximate composition of yoghurt, and viability of starter cultures in yoghurt. Methods: The LAB starter cultures were selected based on their ability to produce diacetyl and lactic acid. Results: Lactobacillus caseiN1 produced the highest quantity (2.72 g/L) of diacetyl at 48 hrs of incubation while Pediococcus acidilacticiG1 had the lowest amount (0.50 g/L). The pH of produced yoghurt ranged between 4.40 and 5.58 while the corresponding lactic acid contents ranged between 0.70 and 0.96 g/L. Yoghurt produced with cow milk inoculated with L. PlantarumN24 and L. BrevisN10 had the lowest pH (4.40) at significant level of P≤0.05. Yoghurt with mixed culture of L. PlantarumN24 and L. PlantarumN17 had the highest protein content (5.13%) while spontaneous fermentation (control) produced the least (0.48%). Yoghurt produced from cow milk inoculated with L. PlantarumN24 and L. PlantarumN17 was rated best with overall acceptability (9.0) during first day of storage while the commercial yoghurt (5.8) and spontaneous fermentation (6.8) had least overall acceptability at P≤0.05. Conclusion: Yoghurt samples stored in refrigerator had more viable LAB counts for a period of 21 days while the samples stored at room temperature had a day count except for yoghurt produced with cow milk inoculated with L. plantarumN24 which retained its viability at the second day. The yoghurt produced with selected LAB starters are better than commercial yoghurt in terms of sensory properties, proximate composition, pH and viability.

Pengaruh Pemberian Bakteri Bacillus pumilus pada kotoran Sapi Sebagai Pupuk terhadap Jumlah Kandungan Klorofil Dunaliella salina [The Effect of Bacteria Bacillus pumilus In Cow Dung As Fertilizer to Total Chlorophyll Dunaliella salina]

Dunaliella salina is one type of natural food used in the hatchery business. Production stability D. salina with an abundance of nutrients can be supported. The purpose of this study was to determine the effect and how many doses of the bacteria Bacillus pumilus best fermented in cow dung as fertilizer in increasing the population of D. salina. Research conducted at the Laboratory of Education Faculty of Fisheries and Marine Fisheries, Airlangga University Surabaya. The research design used was Completely Randomized Design (CRD) is followed by Duncan test. D. salina was cultured in 250 mL glass bottle with 5 treatments (2 treatments as a control) and four replications. Culture medium used contained 10 ppm cow dung. The concentration of cow manure are given in the study, namely A (12.5% Bacillus pumilus), B (10% Bacillus pumilus), C (7.5% Bacillus pumilus), control 1 (without fermentation), control 2 (fertilizer Walne). The results showed that the addition of Bacillus pumilus on cow dung that were cultured in culture medium D. salina can increase the amount of chlorophyll content of D. salina. Addition of Bacillus pumilus by 10% in the culture medium to produce the amount of chlorophyll-a D. salina high of 0.014055 µg/mL and chlorophyll-b of 0.009657142 µg/mL on the first day.

Investigation on Potential Starter of Bacillus spp. for Ivorian Cocoa Beans Fermentation Improvement

To investigate some interesting functional properties of Bacillus spp. in cocoa fermentation processing, 702 strains of Bacillus sp. strains were isolated from six main Ivorian cocoa producer regions. Three mains technological and useful properties for good cocoa beans fermentation were monitored. Among the 386 pectinolytic Bacillus spp. strains, 86.79% of them were confirmed for enzymes production in liquid medium. These 335 pectinolytic Bacillus strains displayed variety of pectinolytic activity with 50.45% of them producing both pectinase and polygalacturonase (PG) enzymes. Among these 169 strains, 70% displayed both the ability to acidify the medium and to degrade the citrate. In mimic conditions of pH and temperature variations as for fermentation heap, 11 strains were able to maintain their activity ranged within 2.12 ± 0.02 to 0.41 ± 0.03 µmol/min/mg of proteins for PG and 2.36 ± 0.03 to 0.25 ± 0.03 µmol/min/mg of proteins for pectinase. Two species (T10T2 and T6HS10) identified as Bacillus subtilus and Bacillus thuringiensis, were selected as the best based on pectinase production level. They were able to continuously produce both pectinolytic enzymes in different conditions of organic acids and ethanol contents. The highlighted properties in this study and the performance of these two strains may make them best candidate as starters for cocoa fermentation control.

Nutrient Signaling via the TORC1-Greatwall-PP2AB55δ Pathway Is Responsible for the High Initial Rates of Alcoholic Fermentation in Sake Yeast Strains of Saccharomyces cerevisiae

ABSTRACT Saccharomyces cerevisiae sake yeast strain Kyokai no. 7 (K7) and its relatives carry a homozygous loss-of-function mutation in the RIM15 gene, which encodes a Greatwall family protein kinase. Disruption of RIM15 in nonsake yeast strains leads to improved alcoholic fermentation, indicating that the defect in Rim15p is associated with the enhanced fermentation performance of sake yeast cells. In order to understand how Rim15p mediates fermentation control, we here focused on target-of-rapamycin protein kinase complex 1 (TORC1) and protein phosphatase 2A with the B55δ regulatory subunit (PP2AB55δ), complexes that are known to act upstream and downstream of Rim15p, respectively. Several lines of evidence, including our previous transcriptomic analysis data, suggested enhanced TORC1 signaling in sake yeast cells during sake fermentation. Fermentation tests of the TORC1-related mutants using a laboratory strain revealed that TORC1 signaling positively regulates the initial fermentation rate in a Rim15p-dependent manner. Deletion of the CDC55 gene, encoding B55δ, abolished the high fermentation performance of Rim15p-deficient laboratory yeast and sake yeast cells, indicating that PP2AB55δ mediates the fermentation control by TORC1 and Rim15p. The TORC1-Greatwall-PP2AB55δ pathway similarly affected the fermentation rate in the fission yeast Schizosaccharomyces pombe, strongly suggesting that the evolutionarily conserved pathway governs alcoholic fermentation in yeasts. It is likely that elevated PP2AB55δ activity accounts for the high fermentation performance of sake yeast cells. Heterozygous loss-of-function mutations in CDC55 found in K7-related sake strains may indicate that the Rim15p-deficient phenotypes are disadvantageous to cell survival. IMPORTANCE The biochemical processes and enzymes responsible for glycolysis and alcoholic fermentation by the yeast S. cerevisiae have long been the subject of scientific research. Nevertheless, the factors determining fermentation performance in vivo are not fully understood. As a result, the industrial breeding of yeast strains has required empirical characterization of fermentation by screening numerous mutants through laborious fermentation tests. To establish a rational and efficient breeding strategy, key regulators of alcoholic fermentation need to be identified. In the present study, we focused on how sake yeast strains of S. cerevisiae have acquired high alcoholic fermentation performance. Our findings provide a rational molecular basis to design yeast strains with optimal fermentation performance for production of alcoholic beverages and bioethanol. In addition, as the evolutionarily conserved TORC1-Greatwall-PP2AB55δ pathway plays a major role in the glycolytic control, our work may contribute to research on carbohydrate metabolism in higher eukaryotes.

PP2AB55δ Responsible for the High Initial Rates of Alcoholic Fermentation in Sake Yeast Strains of Saccharomyces cerevisiae

ABSTRACTSake yeast strain Kyokai no. 7 (K7) and its Saccharomyces cerevisiae relatives carry a homozygous loss-of-function mutation in the RIM15 gene, which encodes a Greatwall-family protein kinase. Disruption of RIM15 in non-sake yeast strains leads to improved alcoholic fermentation, indicating that the defect in Rim15p is associated with the enhanced fermentation performance of sake yeast cells. In order to understand how Rim15p mediates fermentation control, we here focused on target-of-rapamycin protein kinase complex 1 (TORC1) and protein phosphatase 2A with the B55Δ regulatory subunit (PP2AB55δ), complexes that are known to act upstream and downstream of Rim15p, respectively. Several lines of evidence, including our previous transcriptomic analysis data, suggested enhanced TORC1 signaling in sake yeast cells during sake fermentation. Fermentation tests of the TORC1-related mutants using a laboratory strain revealed that TORC1 signaling positively regulates the initial fermentation rate in a Rim15p-dependent manner. Deletion of the CDC55 gene encoding B55δ abolished the high fermentation performance of Rim15p-deficient laboratory yeast and sake yeast cells, indicating that PP2AB55δ mediates the fermentation control by TORC1 and Rim15p. The TORC1-Greatwall-PP2AB55δ pathway similarly affected the fermentation rate in the fission yeast Schizosaccharomyces pombe, strongly suggested that the evolutionarily conserved pathway governs alcoholic fermentation in yeasts. It is likely that elevated PP2AB55δ activity accounts for the high fermentation performance of sake yeast cells. Heterozygous loss-of-function mutations in CDC55 found in K7-related sake strains may indicate that the Rim15p-deficient phenotypes are disadvantageous to cell survival.IMPORTANCEThe biochemical processes and enzymes responsible for glycolysis and alcoholic fermentation by the yeast S. cerevisiae have long been the subject of scientific research. Nevertheless, the factors determining fermentation performance in vivo are not fully understood. As a result, the industrial breeding of yeast strains has required empirical characterization of fermentation by screening numerous mutants through laborious fermentation tests. To establish a rational and efficient breeding strategy, key regulators of alcoholic fermentation need to be identified. In the present study, we focused on how sake yeast strains of S. cerevisiae have acquired high alcoholic fermentation performance. Our findings provide a rational molecular basis to design yeast strains with optimal fermentation performance for production of alcoholic beverages and bioethanol. In addition, as the evolutionarily conserved TORC1-Greatwall-PP2AB55δ pathway plays a major role in the glycolytic control, our work may contribute to research on carbohydrate metabolism in higher eukaryotes.

Physiological Response of Bali Cattle to Feeding Substitution of Balinese Fermentation Straw

This study aims to see the physiological response of Bali cattle to the feeding of fermented straw substitution. A total of 12 head of Bali cattle average age five years. This research was conducted from August to September 2017 in Cenrana Village, Kahu Sub-district, Bone District, which is a location in the business of Berdikari Berdikari Mallomo Utama. This study used a randomized block design (RBD) with 4 treatments and 3 replications so that there were 12 experimental units, the procedure was P0: 100% elephant grass + 100% fermentation control, P1: elephant grass 75% + generation fermentation 25%, P2: 50%, elephant grass + 50% fermenting straw, and P3: Elephant grass 25% + 75% fermentation straw. The parameters observed were body temperature, respiration rate, and liver rate. The data obtained were analyzed variance and continued with LRD test. The results showed that the substitution of hay fermentation had a significant effect (P <.01) on the physiological response of Balinese cow heart, but no significant effect (P> .05) to the body temperature and the rate of respiration of Bali cattle. The conclusion that by giving fermented straw substitution feed provide an effect on heart rate (58.33 / min) with treatment 25% elephant grass + 75% fermented straw in the normal range for Bali cattle.

Teaching process control in food engineering: dynamic simulation of a fermentation control process

Students usually have difficulties to understand abstract concepts of process control. Implementing in teaching process the inquiry-based learning helps students to follow methods and practices similar to those of professional scientists in order to construct knowledge. The paper describes the steps reached in simulation-based learning: from experimental data obtained by the students in their practical method (study and measurement of variables to some fermentation processes) to the simulated the behaviour of the process under a feedback control system. By providing opportunities for students to check their understanding and reflect on their learning process performance is enhanced over a traditional lecture course.