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Use of starter cultures of lactic acid bacteria and yeasts in the preparation of togwa, a Tanzanian fermented food.

Starter cultures of lactic acid bacteria (Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum and Pediococcus pentosaceus) and yeasts (Candida pelliculosa, Candida tropicalis, Issatchenkia orientalis and Saccharomyes cerevisiae) isolated from native togwa were tested singly or in combination for their ability to ferment maize-sorghum gruel to produce togwa. All species of bacteria showed an ability to ferment the gruel as judged by lowering the pH from 5.87 to 3.24-3.49 and increasing the titratable acidity from 0.08% to 0.30-0.44% (w/w, lactic acid) in 24 h. Yeasts used singly showed little activity within 12 h, but lowered the pH to 3.57-4.81 and increased the acidity to 0.11-0.21% in 24 h. Yeasts in co-culture with lactic acid bacteria (LAB) had a modest effect on the final acidity (P<0.05). The number of lactic acid bacteria and yeasts increased while the Enterobacteriaceae decreased with fermentation time. The pH was lowered and lactic acid produced significantly (P<0.05) fastest in natural togwa fermentation and in samples fermented by L. plantarum or L. plantarum in co-culture with I. orientalis. The content of fermentable sugars was reduced during fermentation. Most volatile flavour compounds were produced in samples from fermentation by P. pentosaceus and I. orientalis in co-culture with either L. plantarum or L. brevis.

Carbohydrate Metabolism↗

The microflora of fermented nixtamalized corn.

Nixtamalization is a traditional process that improves the nutritional quality of corn. To provide a means of utilizing the nutritional benefits of nixtamalized corn and improve product acceptability, lactic acid fermentation was applied. The objective of the study was to study the microbial profile and establish the important lactobacilli of fermenting nixtamalized corn dough. Two batches of cleaned whole corn were subjected to the process of nixtamalization, using two concentrations of lime (0.5 or 1.0%), milled, made into a dough (50% moisture) and fermented spontaneously for 72 h. A control sample was prepared without alkaline treatment. pH and titratable acidity of the dough were measured. Aerobic mesophiles, lactic acid bacteria, yeasts and molds were enumerated on Plate Count Agar (PCA), deMan, Rogossa and Sharpe (MRS) Agar and Malt Extract Agar (MEA), respectively. The identity of lactobacilli present was established at the species level using API 50 CHL. The pH of all the fermenting systems decreased with fermentation time with concomitant increase in titratable acidity. Lactic acid bacteria in numbers of 1.6 x 10(9), 2.3 x 10(9) and 1.8 x 10(9) cfu/g, respectively yeasts and molds, and numbers of 8.0 x 10(7), 5.0 x 10(5) and 1.7 x 10(5) cfu/g, respectively were observed in the control and the two nixtamalized (0.5% and 1.0% lime) samples after 48 h of fermentation. Lactobacilli identified in the fermenting nixtamalized corn dough were Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus cellobiosus as well as Pediococcus spp. The study demonstrates that nixtamalized corn though alkaline in nature can be subjected to spontaneous fermentation to produce a sour product.

Colony Count, Microbial↗

Changes in the contents of fat-soluble vitamins and provitamins during tempe fermentation.

The content and formation of fat-soluble vitamins and provitamins during tempe fermentation was studied with 14 different strains of Rhizopus sp. All strains investigated form carotenoids in small amounts during fermentation, but beta-carotene formation was only found in fermentations with six Rhizopus strains. The highest increase of beta-carotene production was to be seen from 34 to 48 h of fermentation. Soybean seeds did not contain ergosterol in detectable amounts. Ergosterol was produced by all Rhizopus strains in concentrations of up to 750 microg g(-1) tempe dry weight (dw) during a 34 h fermentation period and of up to 1610 microg g(-1) tempe dw during a 96 h fermentation period. The amounts of gamma-tocopherol in soybeans ranged from 192.4 to 231.8 microg g(-1) dw. Soybean seeds contained only free, not esterified tocopherols. During fermentation the total amount of vitamin E remained constant but the content of free tocopherols decreased. The content of vitamin K1 in soybeans is not strongly effected by tempe fermentation with pure cultures of Rhizopus sp.

Chromatography, High Pressure Liquid↗

Fermentation of low-salt miso as affected by supplementation with ethanol.

Steam-cooked soybeans and rice koji were combined (1:1, w/w), mixed with 5% (w/w) NaCl and ground into a fine paste. Samples (30 g) were deposited in nylon/polyethylene plastic bags and supplemented with 10 ml of aqueous ethanol solutions to give concentrations of 0, 2.5, 5, 7.5, 10, 15, 20, and 25% ethanol. Mixtures were homogenized, sealed, and incubated at 28 degrees C for eight weeks. Mold populations were less than 3 log10 CFU/g in all miso products after four weeks of fermentation. Yeast populations increased to 6.1 log10 CFU/g in the control (0% added ethanol) during the first week of fermentation and remained stable throughout the eight-week fermentation period. Yeasts were not detected in products containing 5-25% ethanol. Populations of lactic acid bacteria (LAB) increased to 6 log10 CFU/g after one week of fermentation in products containing 0 and 2.5% ethanol. However, after eight weeks of fermentation, LAB populations in all products were less than 4 log10 CFU/g. Rapid decreases in pH occurred only in products supplemented with 0 or 2.5% ethanol. Percentages of soluble protein in miso products containing various ethanol concentrations during the eight-week fermentation period revealed that protease activity was still active or not greatly inhibited in products supplemented with less than 10% ethanol. In comparison, koji enzymes were comparatively less affected by ethanol than were populations of molds, yeasts, and LAB. Total soluble carbohydrate and glucose contents were higher in products supplemented with 5, 7.5 and 10% ethanol than in other products. Discoloration (browning) during fermentation occurred most rapidly in products supplemented with 5 or 7.5% ethanol. Sensory evaluation of the low-salt (5%) product supplemented with 7.5% ethanol and fermented for eight weeks revealed normal or enhanced flavor ratings compared to ratings for a commercial product.

Aspergillus oryzae↗

Food fermentations: role of microorganisms in food production and preservation.

Preservation of foods by fermentation is a widely practiced and ancient technology. Fermentation ensures not only increased shelf life and microbiological safety of a food but also may also make some foods more digestible and in the case of cassava fermentation reduces toxicity of the substrate. Lactic acid bacteria because of their unique metabolic characteristics are involved in many fermentation processes of milk, meats, cereals and vegetables. Although many fermentations are traditionally dependent on inoculation from a previous batch starter cultures are available for many commercial processes such as cheese manufacture thus ensuring consistency of process and product quality. This review outlines the role of lactic acid bacteria in many such fermentations and the mechanisms of antibiosis with particular reference to bacteriocins and gives a brief description of some important fermented foods from various countries. It is anticipated that the contribution of the advances in lactic acid bacteria research towards improvement of strains for use in food fermentation will benefit both the consumer and the producer.

Fermentation↗

Potential use of traditional fermented foods for weaning in Zimbabwe.

The current interest in the use of traditional fermented and malted foods for weaning is their potential to reduce the transmission of bacterial enteric pathogens through contaminated weaning foods as well as improving the nutritional value of the foods. This study was carried out to investigate the potential use of traditional fermented and malted foods for weaning in rural areas. Information was obtained by interviewing randomly selected 150 rural women with children under five years old. Ninety seven percent of the women had knowledge of some traditional fermented foods. The commonest fermented foods known by the women were mahewu (traditional fermented and malted sour, non-alcoholic cereal gruel), sour milk and sour porridge. Most of the women indicated that mahewu (94%), sour milk, (87%) and sour porridge (71%) were consumed by all family members. The majority of the women gave the fermented foods to infants from the age of four months. Although most of the infants were introduced to the fermented and malted foods from the age of four months, the foods were not given to infants very often, with the exception of mahewu. The storage period of the fermented foods ranged from one to three days. The results of this study show that mahewu, sour milk and sour porridge have a potential for use as weaning foods if they are promoted.

Child Nutrition Sciences↗

Prevention of gastrointestinal infection using immunobiological methods with milk fermented with Lactobacillus casei and Lactobacillus acidophilus.

The protective effect of feeding milk fermented with a mixture of Lactobacillus casei sp. and Lb. acidophilus sp. against Salmonella typhimurium infection in mice was compared with that obtained feeding milks fermented with these microorganisms individually. The survival rate obtained after oral infection with Sal. typhimurium was 100% in mice pretreated by feeding during 8 d with the mixture of Lb. casei and Lb. acidophilus fermented milks. Similar treatments with the individual milks were ineffective. Moreover, mice became more susceptible to infection with Sal. typhimurium after such treatment. The colonization of liver and spleen with the pathogen was markedly inhibited by the pretreatment with the mixture of fermented milk, while such inhibition was not observed using the Lb. casei and Lb. acidophilus milks. The highest levels of anti-salmonellae antibodies in serum and in intestinal fluid were found in the group of mice fed with the mixture and with Lb. casei fermented milk respectively. However, this latter milk was not effective in protecting against Sal. typhimurium. When the mice were first infected with Sal. typhimurium and then fed with the mixture of fermented milks, pathogen colonization was not prevented. The results suggest that the augmentation of resistance to salmonellae caused by the treatment with Lb. casei- + Lb. acidophilus-fermented milk was due to the anti-salmonellae protective immunity mainly mediated by the mucosal tissue. Milk fermented with this mixture could be used as an immunobiological method to prevent gastrointestinal infection.

Animals↗

Measurement and analysis of intracellular ATP levels in metabolically engineered Zymomonas mobilis fermenting glucose and xylose mixtures.

Intracellular adenosine-5'-triphosphate (ATP) levels were measured in a metabolically engineered Zymomonas mobilis over the course of batch fermentations of glucose and xylose mixtures. Fermentations were conducted over a range of pH (5-6) in the presence of varying initial amounts of acetic acid (0-8 g/L) using a 10% (w/v) total sugar concentration (glucose only, xylose only, or 5% glucose/5% xylose mixture). Over the design space investigated, ethanol process yields varied between 56.6% and 92.3% +/- 1.3% of theoretical, depending upon the test conditions. The large variation in process yields reflects the strong effect pH plays in modulating the inhibitory effect of acetic acid on fermentation performance. A corresponding effect was observed on maximum cellular specific growth rates, with the rates varying between a low of 0.15 h(-1) observed at pH 5 in the presence of 8 g/L acetic acid to a high of 0.32 +/- 0.02 h(-1) obtained at pH 5 or 6 when no acetic acid was initially present. While substantial differences were observed in intracellular specific ATP concentration profiles depending upon fermentation conditions, maximum intracellular ATP accumulation levels varied within a relatively narrow range (1.5-3.8 mg ATP/g dry cell mass). Xylose fermentations produced and accumulated ATP at much slower rates than mixed sugar fermentations (5% glucose, 5% xylose), and the ATP production and accumulation rates in the mixed sugar fermentations were slightly slower than in glucose fermentations. Results demonstrate that higher levels of acetic acid delay the onset and influence the extent of intracellular ATP accumulation. ATP production and accumulation rates were most sensitive to acetic acid at lower values of pH.

Adenosine Triphosphate↗

Plant-derived biomolecules in fermented cabbage.

The formation of plant-derived biomolecules during sauerkraut fermentation was studied. Cabbage was fermented with a starter culture, and the results were compared to the results of spontaneous fermentation. The concentration of flavonoids and glucosinolates was analyzed by HPLC, and that of the glucosinolate breakdown products, by GC-MS. Of the 20 different flavonoids tested, only kaempferol was found (0.9 mg/ kg FW, fresh weight). The content of kaempferol remained constant in the cabbage fiber matrix over the fermentation process. The nitrite concentration was below the detection limit in both fermentations. The total glucosinolate content in the raw material was 3.71 micro mol/g DW, dry weight. Glucosinolates were totally decomposed in both fermentations during two weeks, and different types of breakdown products were formed. Isothiocyanates, indole-3-carbinol, goitrin, allyl cyanide, and nitriles were determined in the fermented cabbage. Isothiocyanates and allyl cyanide were the predominant breakdown products in both fermentations. Sulforaphane nitrile and goitrin were found only in small quantities in the end products.

Brassica↗

Antioxidant activity of fermented soybean extract.

Free radicals are considered to be important causative factors in the development of cancer and cardiovascular diseases. This relationship has led to interest in evaluating the antioxidant capacities of many dietary supplements. Fermented soybean extract is produced by symbiotic fermentation of organic soybean with 20 types of Lactobacillus and yeast. In vitro and in vivo models are used in this study to evaluate the antioxidant effect of fermented soybean extract. Several in vitro models are used to detect the antioxidant capacity of the fermented soybean extract, which is compared to vitamin C and Trolox. The results demonstrate that the fermented soybean extract has strong antioxidant activity against unsaturated fatty acid peroxidation compared to vitamin C and Trolox. By the means of the test system developed by Y. Toshiki et al., it is shown that the fermented soybean extract can function both as an antioxidant and as a free radical acceptor that can convert free radicals into harmless substances through an energy-decreasing procedure. An in vivo study examines the effects of fermented soybean extract on the activity of antioxidant enzymes. The activities of the antioxidant enzymes (AOE) including total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) are measured in liver, kidney, and brain from male Sprague-Dawley rats. The activities of CAT, SOD, and GPX are increased in the liver. However, the SOD activity is decreased in the kidney. SOD and GPX activities are decreased in the brain. These results lead to the conclusion that fermented soybean extract not only has antioxidant activity but also has an effect on the activity of antioxidant enzymes in liver.

Animals↗

Volatile constituents of fermented sugar baits and their attraction to lepidopteran species.

The volatile compounds emanating from four fermented sugar baits, palm sugar, golden cane syrup, port wine, and molasses, were isolated by headspace sampling and analyzed by gas chromatography-mass spectrometry. Three classes of compounds including esters, alcohols, and aromatic compounds were identified in the headspace of the four fermented sugar baits. There was a high degree of qualitative similarity between the headspace contents of the four fermented sugar baits, although quantitatively they varied considerably. Ethyl acetate, 3-methylbutanol, ethyl hexanoate, 2-phenylethanol, ethyl octanoate, ethyl (E)-4-decenoate, ethyl decanoate, and ethyl dodecanoate were the major compounds identified in the headspace of the four fermented sugar baits. The efficacy of the four fermented sugar baits was investigated in field trapping experiments. Fermented palm sugar and golden cane syrup were superior in attracting significant numbers of moths as compared to port wine and molasses. Fermented molasses was the least attractive among the four baits. Over 90% of the insects caught were noctuids with Graphania mutans and Tmetolophota spp. being the main noctuids captured (over 55%) in the four fermented sugar baits. Male and female G. mutans were equally attracted to the four sugar baits. A number of tortricid species were also trapped.

Alcohols↗

Study on furundu, a traditional Sudanese fermented roselle (Hibiscus sabdariffa L.) seed: effect on in vitro protein digestibility, chemical composition, and functional properties of the total proteins.

Furundu, a meat substitute, is traditionally prepared by cooking the karkade (Hibiscus sabdariffa L.) seed and then fermenting it for 9 days. Physicochemical and functional properties of raw and cooked seed and of furundu ferments were analyzed. Furundu preparation resulted in significant changes in karkade seed major nutrients. Total polyphenols and phytic acid were also reduced. The increase in total acidity and fat acidity coupled with a decrease in pH indicates microbial hydrolysis of the major nutrients; proteins, carbohydrates, and fats. In vitro digestibility of the seed proteins reached the maximum value (82.7%) at the sixth day of fermentation, but thereafter it significantly decreased. The effect of furundu preparation on N solubility profiles and functional properties, such as emulsification and foaming properties and other related parameters, is investigated in water and in 1 M NaCl extracts from defatted flour samples. The results show that cooking followed by fermentation affects proteins solubility in water and 1 M NaCl. The foaming capacity (FC) from the flour of raw seed decreased as a result of cooking. Fermentation for 9 days significantly increased the FC of the cooked seed, restoring the inherent value. Foam from fermented samples collapsed more rapidly during a period of 120 min as compared to the foam from raw and cooked karkade seeds; stability in 1 M NaCl was lower as compared to those in water. In water, the emulsion stability (ES) from the fermented samples was significantly higher than that of the raw seed flour. Addition of 1 M NaCl significantly decreased the ES of the fermented samples.

Chemical Phenomena↗

Fermented pigeon pea (Cajanus cajan) ingredients in pasta products.

Pigeon pea (Cajanus cajan var. aroíto) seeds were fermented in order to remove antinutritional factors and to obtain functional legume flour to be used as pasta ingredients. Fermentation brought about a drastic reduction of alpha-galactosides (82%), phytic acid (48%), and trypsin inhibitor activity (39%). Fermented legume flours presented a notable increase of fat and total soluble available carbohydrates, a slight decrease of protein, dietary fiber, calcium, vitamin B2, vitamin E, and total antioxidant capacity, and a decrease of soluble dietary fiber, Na, K, Mg, and Zn contents. No changes were observed in the level of starch and tannins as a consequence of fermentation. The fermented flour was used as an ingredient to make pasta products in a proportion of 5, 10, and 12%. The supplemented pasta products obtained had longer cooking times, higher cooking water absorptions, higher cooking loss, and higher protein loss in water than control pasta (100% semolina). From sensory evaluations, fortified pasta with 5 and 10% fermented pigeon pea flour had an acceptability score similar to control pasta. Pasta supplemented with 10% fermented pigeon pea flour presented higher levels of protein, fat, dietary fiber, mineral, vitamin E, and Trolox equivalent antioxidant capacity than 100% semolina pasta and similar vitamins B1 and B2 contents. Protein efficiency ratios and true protein digestibility improved (73 and 6%, respectively) after supplementation with 10% fermented pigeon pea flour; therefore, the nutritional value was enhanced.

Cajanus↗

Influence of temperature on the fermentation of bambara groundnut (Vigna subterranea) to produce a dawadawa-type product.

Bambara groundnut (Vigna subterranea) was fermented to produce a dawadawa-type product using a starter culture of Bacillus licheniformis isolated from naturally fermenting bambara groundnut beans. Fermentation was carried out at 30 and 37 degrees C for four days and at 45 degrees C for two days. The pH of the substrate decreased after 24 hours and then rose at 30 and 37 degrees C but remained constant at 45 degrees C after the initial drop. Total titratable acidity of the fermenting beans mimicked the pH values. Proximate analyses for moisture, protein and fat of the cotyledons showed an increase in all three constituent at each of the three fermentation temperatures. At the end of fermentation, total available carbohydrate was 55%, 59% and 62% of the original value at 30, 37 and 45 degrees C, respectively. Fermentation of bambara groundnut at 45 degrees C for two days is recommended as the ideal fermentation temperature and time.

Africa↗

Effects of resistant starch, a non-digestible fermentable fiber, on reducing body fat.

OBJECTIVE: To assess the effects of energy dilution with non-fermentable and fermentable fibers on abdominal fat and gut peptide YY (PYY) and glucagon-like peptide (GLP)-1 expressions, three rat studies were conducted to: determine the effects of energy dilution with a non-fermentable fiber, compare similar fiber levels of fermentable and non-fermentable fibers, and compare similar metabolizable energy dilutions with fermentable and non-fermentable fibers. RESEARCH METHODS AND PROCEDURES: In Study 1, rats were fed one of three diets with different metabolizable energy densities. In Study 2, rats were fed diets with similar fiber levels using high amylose-resistant cornstarch (RS) or methylcellulose. In Study 3, rats were fed diets with a similar dilution of metabolizable energy using cellulose or RS. Measurements included food intake, body weight, abdominal fat, plasma PYY and GLP-1, gastrointestinal tract weights, and gene transcription of PYY and proglucagon. RESULTS: Energy dilution resulted in decreased abdominal fat in all studies. In Study 2, rats fed fermentable RS had increased cecal weights and plasma PYY and GLP-1, and increased gene transcription of PYY and proglucagon. In Study 3, RS-fed rats had increased short-chain fatty acids in cecal contents, plasma PYY (GLP-1 not measured), and gene transcription for PYY and proglucagon. DISCUSSION: Inclusion of RS in the diet may affect energy balance through its effect as a fiber or a stimulator of PYY and GLP-1 expression. Increasing gut hormone signaling with a bioactive functional food such as RS may be an effective natural approach to the treatment of obesity.

Adipose Tissue↗

The effect of consumption of milk fermented by Lactobacillus casei strain Shirota on the intestinal microflora and immune parameters in humans.

OBJECTIVE: To determine the effect of consumption of milk fermented by Lactobacillus casei strain Shirota (L. casei Shirota) on the composition and metabolic activities of the intestinal microflora, and immune parameters in humans. SUBJECTS: Twenty healthy male subjects aged 40-65 years were selected. DESIGN: A placebo-controlled trial was performed in which 10 subjects were randomly assigned to a control and 10 to a treatment group. During the first and last two weeks of the 8-week study the subjects received a strictly controlled diet without fermented products. The same controlled diet was given during the intermediate 4-week test period but then the treatment group received three times daily 100 ml of fermented milk containing 10(9) CFU L. casei Shirota/ml, whereas the same amount of unfermented milk was given to the subjects in the control group. RESULTS: In comparison to the control group, the consumption of L. casei Shirota-fermented milk resulted in an increase of the Lactobacillus count in the faeces in which the administered L. casei Shirota was predominant at the level of 10(7) CFU/g wet faeces. This was associated with a significant increase in Bifidobacterium counts (P < 0.05). Some shifts in the other bacterial species were found, such as a decreased number of Clostridium; however the differences were not statistically different between the treatment and the control groups. The beta-glucuronidase and beta-glucosidase activities per 10(10) bacteria decreased significantly (P < 0.05) at the second week of the 4-week test period with the consumption of L. casei Shirota-fermented milk. Furthermore, the consumption of the fermented milk product resulted in a slight but significant increase in the moisture content of the faecal samples (P < 0.05). No treatment effects were observed for any of the immune parameters measured (including natural killer (NK) cell activity, phagocytosis and cytokine production). CONCLUSIONS: The results suggest that consumption of L. casei Shirota-fermented milk is able to modulate the composition and metabolic activity of the intestinal flora and indicate that L. casei Shirota-fermented milk does not influence the immune system of healthy immunocompetent males.

Adult↗

Microbial colonization of naturally black olives during fermentation and associated biochemical activities in the cover brine.

AIMS: To establish the site of microbial growth on naturally black fermented table olives, and to monitor the population dynamics of yeasts and selected micro-organisms together with the changes in organic acid profile and pH in the cover brine during fermentation. METHODS AND RESULTS: During fermentation, the numbers of Enterobacteriaceae and Pseudomonas spp. in the brine decreased whilst lactic acid bacteria and yeast populations increased. Scanning electron microscopy showed that a yeast-rich biofilm developed on the epicuticular wax of the olive skin during fermentation. Yeasts also predominated in the stomatal openings, but bacteria were more numerous in intercellular spaces in the sub-stomatal flesh. Citric, malic and tartaric acids were the major organic acids accumulating in the brine during fermentation. CONCLUSIONS: Micro-organisms associated with the skin, stomata and flesh in fermenting black olives may experience different local conditions to those prevailing in the cover brine. SIGNIFICANCE AND IMPACT OF THE STUDY: These are the first observations of the micro-organisms associated with the fruit of naturally fermented black olives and of the accumulation of specific organic acids during fermentation.

Biofilms↗

The use of cumulative gas and volatile fatty acid production to predict in vitro fermentation kinetics of Italian ryegrass leaf cell walls and contents at various time intervals.

Differences between the fermentation characteristics of cell contents (CC) and protease-treated cell walls (CW) of young leaves of Italian ryegrass (Lolium multiflorum Lam.) cultivar Multimo (tetraploid), were studied in vitro. Gas and volatile fatty acid (VFA) production rates were measured at regular intervals, as was the degradability of organic matter (OM) of CW. The measured VFA were used to predict the gas production and fermentable OM using stoichiometric calculations. For CW the volume and kinetics of measured gas production were the same as those predicted from the VFA formed. In contrast, the measured gas production for CC was consistently less than predicted, indicating that the stoichiometric equations were not valid for rapidly fermenting substrates. For both CC and CW, the relative rate of acetic acid production levelled off more slowly than for other VFA, resulting in an increasing gas yield (in ml/g fermentable OM) after 12 (CW)-24 (CC) h incubation. Consequently, the fermentation of OM was not linearly related to gas production kinetics. For CW, the kinetics of decline of degradable OM and fermentable OM were the same, after correction for a constant 'lost fraction' of degradable OM of 205 g/kg OM. This work indicates the value of detailed studies of fermentation processes to evaluate herbage quality. In particular, the role of CC and the difference between degradation and fermentation require further attention.

Cell Wall↗