Heavy metal contents and chemical compositions of atlantic (<Emphasis Type="Italic">Scomber scombrus</Emphasis>), blue (<Emphasis Type="Italic">Scomber australasicus</Emphasis>), and chub (<Emphasis Type="Italic">Scomber japonicus</Emphasis>) mackerel muscles

The heavy metal contents and chemical compositions among mackerel species were investigated. Atlantic and chub mackerel had higher mercury, but lower lead than blue mackerel. Chub mackerel had the highest crude fat (18.62%) and the lowest moisture (58.29%), whereas blue mackerel had the highest moisture (70.10%) and the lowest crude fat (2.89%). Atlantic and blue mackerel had total saturated fatty acids (SFA)>polyunsaturated fatty acids (PUFA)>monounsaturated fatty acids (MUFA), whereas chub mackerel had PUFA>SFA>MUFA. The main SFA of the 3 mackerels were 16:0 and the main PUFAs were 22:6n-3 and 20:5n-3. The total amino acid contents in Atlantic, blue, and chub mackerel were 190.63, 169.49, and 172.97 mg/g, respectively. The major amino acids of the 3 mackerels were glutamic acid, aspartic acid, lysine, and leucine.     

Estrogenic effects of various extracts from <Emphasis Type="Italic">Chamdanggui</Emphasis> (<Emphasis Type="Italic">Angelica gigas</Emphasis> Nakai) and <Emphasis Type="Italic">sogdan</Emphasis> (<Emphasis Type="Italic">Phlomis umbrosa</Emphasis> Turcz)

The various extracts from chamdanggui (Angelica gigas Nakai) and sogdan (Phlomis umbrosa Turcz) were evaluated for estrogenic activity and characterized according to HPLC profile. Chamdanggui and sogdan were individually extracted with 4 solvents (hot water, 70% ethanol, n-butanol, and dichloromethane) of differing polarities. Estrogenic activity was determined by E-screen using an estrogen-dependent MCF-7 BUS cell. Although almost all extracts showed estrogenic effects in a concentrationdependent manner, the hot water extract from chamdanggui (250 μg/mL) had the higher effect (138%). Among 90 fractions using HPLC separation of the hot water extract from chamdanggui, fraction 21 and 28 produced the highest estrogenic effects of 178 and 163% at 10 μg/mL, respectively. The results imply that the hot water extract from chamdanggui could be useful as an alternative hormone replacement therapy.     

Combined Effect of Icing Medium and Red Alga <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis> on Shelf Life Extension of Indian Mackerel (<Emphasis Type="Italic">Rastrelliger kanagurta</Emphasis>)

Flake ice refrigeration is a conventional but commonly employed technique to slow down fish spoilage and deterioration of nutritional values and sensory properties during chilled storage. In the present study, a methanolic extract of red alga Gracilaria verrucosa was characterised, and functional groups, such as alkenes, aldehydes, nitriles, galactans and galactose-4-sulphate were found. Subsequent identification of active compounds revealed the presence of potent preservative agents, such as butylated hydroxytoluene, sulfurous acid, 1,2-propanediol, benzeneacetic acid, cyclononasiloxane and tetracosamethyl-cyclododecasiloxane. The effect of incorporating G. verrucosa at two different concentrations (0.67 and 2.5 g lyophilised alga/L aqueous solution) in the icing medium was tested for the preservation of Indian mackerel (Rastrelliger kanagurta), compared to preservation in traditional ice prepared only from water. Microbial, chemical and sensory qualities were monitored in Indian mackerel chilled in ice with and without G. verrucosa during 15 days storage period. Inhibitory effects (p?<?0.05) on the microbial proliferation (mesophilic and psychrophilic bacteria) and chemical markers of fish deterioration (pH, TVB-N, TMA-N and biogenic amines) were evidenced for ice containing both concentrations of G. verrucosa, respectively, relative to the control medium. The sensory score acceptability limit reached 11 days for Indian mackerel stored in traditional ice and 15 days for Indian mackerel stored in ice with G. verrucosa extract. Thus, the icing medium containing G. verrucosa extract improves the quality and safety of Indian mackerel during storage and can be explored by the seafood industry as a biopreservative.     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Thermal inactivation kinetics of quality-related enzymes in cauliflower (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">botrytis</Emphasis><Emphasis Type="Bold">)</Emphasis>

Thermal inactivation of quality-related enzymes in both cauliflower crude enzyme extracts and fresh tissue samples was studied in temperature range 50–100 °C. For crude enzyme extracts, several parameters, reaction rate constants (k) and activation energy (E _a) as well as decimal reduction time (D) and (z) values, were used to characterize the thermal stability. The rates of inactivation were found to follow first-order inactivation kinetics. Activation energies varied between 101.18 and 208.42 kJ mol⁻¹ with z values of 10.59–24.09 °C. The examined kinetics indicated that lipoxygenase was the most heat resistant followed by peroxidase, polyphenol oxidase, pectin methyl esterase and ascorbic acid oxidase. Furthermore, the obtained results from the blanched fresh tissues indicated that inactivation of lipoxygenase secured disappearing of any other enzyme activities. Therefore, this study recommends using lipoxygenase as an indicator enzyme to optimize the thermal treatments of cauliflower products.     

Molecular diagnostics of Lemon Myrtle (<Emphasis Type="Italic">Backhousia citriodora</Emphasis> versus <Emphasis Type="Italic">Leptospermum citratum</Emphasis>)

‘Lemon Myrtle’ is becoming increasingly popular in Europe both for use in cuisine and phytotherapy. However, this common name covers two completely different species, Backhousia citriodora F. Muell. and Leptospermum citratum Challinor, Cheel & A.R.Penfold. These species differ with respect to secondary compounds and even can cause, if mixed up and applied in high dose, toxic effects. We describe how the two species can be discriminated microscopically making use of differences in the morphology of leaf pavement cells and the relative size of palisade parenchyma. Based on the large subunit of ribulose-1,5-bisphosphate carboxylase oxygenase (rbcL) as molecular marker, the phylogenetic position of the two species within the Myrtaceae could be clarified. This sequence information was used to develop a simple assay to discriminate the two species even in dried and highly fragmented mixtures as typically occurring in commercial samples. This assay utilises the occurrence of single-nucleotide exchanges between those species that produce different fragments when the rbcL amplificates are restricted with Sac II.     

Effect of glasswort (<Emphasis Type="Italic">Salicornia herbacea</Emphasis> L.) and rice (<Emphasis Type="Italic">oryza sativa</Emphasis> L.) on quality of <Emphasis Type="Italic">Ganjang</Emphasis> (Korean Soy Sauce)

Four types of meju were made from 100%(w/w) defatted soybean (DFS), a mixture of 80%(w/w) defatted soybean, and 20%(w/w) glasswort (DFS-G), a mixture of 80%(w/w) defatted soybean and 20%(w/w) rice (DFS-R), and a mixture of 60%(w/w) defatted soybean, 20%(w/w) glasswort, and 20%(w/w) rice (DFS-GR). Four types of Korean traditional soy sauce were prepared from the 4 types of meju. Mineral and antioxidant contents in the soy sauce made of DFS-G and DFS-GR were significantly higher than others. Citric, malic, succinic, lactic, and pyroglutamic acid contents in soy sauce made of DFS-R and DFS-GR were 1.3–1.5 times higher than others. Total nitrogen and free amino acid contents in soy sauce were correlated with DFS concentration in the meju. The bacterial community in the non-fermented meju-making ingredients was replaced largely by Bacillus sp. in the fermented meju. The use of glasswort and rice in the meju-making process did not alter the bacterial community responsible for the fermentation of meju.     

Drying kinetics of whole and sliced <Emphasis Type="Italic">shiitake</Emphasis> mushrooms (<Emphasis Type="Italic">Lentinus edodes</Emphasis>)

Isotherms of shiitake mushroom (Lentinus edodes) at 25 and 40°C were determined and drying kinetics of whole and sliced shiitake mushrooms were tested using a convective air drying method at different drying temperature of 40, 50, 60, and 70°C. The monolayer moisture contents of the mushroom were 7.23 and 5.44 g water/100 g of dry solids at 25 and 40°C, respectively. Both mushroom samples showed falling drying rate periods with increasing drying rates with increases in drying temperature, and the drying rate of the sliced mushrooms was faster than that of the whole mushrooms at the same drying conditions. The kinetic parameters for dehydration of the mushrooms were determined using the Newton model and the Classical diffusion model. Activation energy (E _a ) values as determined using the Newton model were 22.58 and 20.48 kJ/mol for the whole and sliced mushrooms, respectively.     

Antioxidative activity of<Emphasis Type="Italic"> Geranium macrorrhizum</Emphasis>

The composition of radical-scavenging compounds from Geranium macrorrhizum leaves was analyzed and the antioxidative activities of various extracts was determined. Seven compounds, namely gallic acid, ellagic acid, 4-galloyl quinic acid, the flavonoid quercetin and three of its glycosides, quercetin-3--glucopyranoside, quercetin-3--galactopyranoside and quercetin-4--glucopyranoside were isolated and identified in the various fractions. The radical-scavenging activity of the isolated compounds was measured using DPPH^· and ABTS^·+ scavenging assays and compared with the activity of rosmarinic acid. Quercetin-3-glucopyranoside and quercetin-3-galactopyranoside showed the highest antioxidative capacities. Antioxidative activities of all plant fractions were assessed by model system tests (superoxide anion and hydrogen peroxide scavenging) and by oxidation tests (-carotene bleaching, peroxide value, UV absorbance and headspace-gas chromatography hexanal determination). The ethanol-butanol fraction of the plant possessed the highest activity in most of the tests.     

Effect of heat-treated <Emphasis Type="Italic">Nuruk</Emphasis> on the quality characteristics of aged <Emphasis Type="Italic">Yakju</Emphasis>

Long-term aging of Yakju, a traditional Korean liquor made of rice and Nuruk (a fermentation agent), causes browning and odor and flavor development. This study investigated the effects of heat-treated Nuruk (50–80 °C, 30 min) on Yakju quality. The saccharogenic powers and glucoamylase, α-amylase, and carboxypeptidase activities were similar in non-heat-treated Nuruk and that treated at 50 °C. However, acidic protease and alcohol dehydrogenase decreased above 50 °C. The content of nitrogen-containing compounds was inversely proportional to the heat-treatment temperature. Compounds that cause off-flavors decreased at 50–60 °C, but increased at 70–80 °C, whereas compounds that provide fragrance increased at 50–60 °C. Sensory evaluation indicated that bad taste attributes were higher in Yakju produced using non-heat-treated Nuruk. Therefore, heat treatment of Nuruk at 50 °C can be adopted as a method for improving Yakju quality, as enzymatic activities that affect color, aroma, and taste are regulated.     

A New Multiplexed Real-Time PCR Assay to Detect <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>, <Emphasis Type="Italic">C. coli</Emphasis>, <Emphasis Type="Italic">C. lari</Emphasis>, and <Emphasis Type="Italic">C. upsaliensis</Emphasis>

A new rapid method based on real-time PCR was developed to detect four thermophilic Campylobacter species (Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis) in food samples. The assay targeted the bipA gene for C. upsaliensis and C. lari, whereas the gene encoding the ATP-binding protein CJE0832 was used to detect C. coli and C. jejuni. These genes were chosen for this assay due to their low variability and mutation rate at a species level. The multiplex PCR showed 100% inclusivity for all 25 thermophilic Campylobacter strains tested and 100% exclusivity for 38 non-targeted strains belonging to closely related species. The newly developed real-time PCR could detect down to 10² genomes/reaction and displayed efficiency above 97% for all species except for C. upsaliensis (90.1%). The method proved to be a reliable tool for food analysis, showing 100% sensitivity, 96% efficiency, and 92.45% specificity when validated against the gold standard method UNE-EN ISO 10272:2006 using 200 diverse food samples (meat, fish, fruits and vegetables, and raw milk). In artificially spiked samples, the detection limit of the method was 10 cfu/g in salad, 5 cfu/g in turkey meat, and 1 cfu/g in the rest of meat samples tested. Consequently, the newly designed molecular tool represents a quick and safe alternative to obtain reliable results concerning the presence/absence of the main thermophilic Campylobacter in any food sample.     

Effect of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> Bauer and <Emphasis Type="Italic">Bifidobacterium animalis</Emphasis> ssp. <Emphasis Type="Italic">lactis</Emphasis> BB12 on proteolytic changes in dry-cured loins

The effect of the potentially probiotic bacteria strain of Lactobacillus acidophilus Bauer and probiotic bacteria Bifidobacterium animalis ssp. lactis BB12 on proteolytic changes of proteins in dry-cured loins during fermentation and cold storage was studied. Results of the conducted tests demonstrated that the use of probiotic bacteria for the production of dry-cured meats impacts the generation of products of protein proteolysis with high antioxidant activity. The highest antioxidant activity of peptides after fermentation and cold storage was observed in the loin with the strain B. animalis ssp. lactis BB12 and the loin with the mixture of strains L. acidophilus Bauer and B. animalis ssp. lactis BB12. Qualitative analysis of peptides demonstrated that peptides with weight below 3.5 kDa are characterized by the highest capacity of quenching the ABTS cation radical, including the peptides in loins with the strain B. animalis ssp. lactis BB12.     

Phenolic contents and antioxidant activities from different tissues of <Emphasis Type="Italic">Baekseohyang</Emphasis> (<Emphasis Type="Italic">Daphne kiusiana</Emphasis>)

The aim of this research was to investigate the levels of phenolic compounds and antioxidant activities in different tissues including leaves, stems, and roots from baekseohyang (Daphne kiusiana). The highest contents of total phenolics (43.59 mg gallic acid equivalent, GAE/g) and flavonoids (15.73 mg rutin equivalents, RE/g) were observed in the 75% methanol extract of leaves. Moreover, this extract had the predominant antioxidant capacity, DPPH (85.91%) and ABTS (92.57%) radical scavenging activities as well as reducing power (7.20%) at a concentration of 5 mg/mL. The highest content of phenolic compounds was also exhibited in this extract with an increasing order in leaves, roots, and stems and their major components were vanillic acid (6.37 mg/g), tannic acid (1.91 mg/g), and p-hydroxybenzoic acid (3.96 mg/g). Thus, the strong antioxidant activities of the 75% methanol extract are correlated with high phenolic compound contents. This study suggests that baekseohyang leaves may potentially be used as an accessible source of natural antioxidants.     

In vivo study of antiallergenicity of ethanol extracts from <Emphasis Type="Italic">Sargassum tenerrimum</Emphasis>, <Emphasis Type="Italic">Sargassum cervicorne</Emphasis> and <Emphasis Type="Italic">Sargassum graminifolium turn</Emphasis>

Food allergy has becoming the serious threat in the world for which the search of an effective anti-allergic drug is the demand of time. Keeping in view of the potentiality of seaweeds, the ethanol extracts from Sargassum tenerrimum (ST), Sargassum cervicorne (SC), and Sargassum graminifolium turn (SG) have been studied in vivo for its antiallergenicity through passive cutaneous anaphylaxis (PCA) and active cutaneous anaphylaxis (ACA) in female BALB/c mice. Intraperitoneal administration of these ethanol extracts inhibit mouse PCA and ACA in a dose-dependent manner using ovalbumin (OVA) and shrimp allergen as triggering agents to induce allergenicity over mice. The extract of ST containing phlorotannin has been found most active over the suppression of PCA triggered by OVA and shrimp with IC₅₀ values of 25.64 and 40.98 mg/kg, respectively and an efficacy comparable to that of an anti-allergic drug disodiumcromoglycate. Similarly, ST inhibits ACA triggered by ova and shrimp allergen in the mouse, with 50% suppression at 25.5 and 43.53 mg/kg, respectively. The results presented here show that these extracts are active on the studied models among which ethanol extract of ST was the most potent, leading toward the promising development of a new class of anti-allergic drugs.     

Molecular diversity among natural populations of <Emphasis Type="Italic">Lactobacillus paracasei</Emphasis> and <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis>/<Emphasis Type="Italic">paraplantarum</Emphasis> strains isolated from autochthonous dairy products

The diversity of 87 Lactobacillus paracasei and Lactobacillus plantarum/paraplantarum strains, previously identified from different autochthonous dairy products, was investigated by phenotypic and genotypic approaches. The increased resolution obtained using phenotypic and genotypic characterization allowed the level of strain heterogeneity detection to be widened. Phenotypic diversity was evaluated by studying biochemical characteristics of technological interest, including antimicrobial and proteinase activities, resistance to nisin, aggregation ability, production of exopolysaccharides, acetoin and diacetyl, citrate utilization, and antibiotic susceptibility. Genotypic diversity was generally evaluated by PCR amplification of repetitive bacterial DNA element fingerprinting using the (GTG)₅ primer [(GTG)₅-PCR]. Moreover, in cases where strains were not discriminated by (GTG)₅-PCR combined with phenotypic analysis, pulsed-field gel electrophoresis (PFGE) analysis was performed. The results indicate that L. plantarum/paraplantarum and L. paracasei natural isolates from artisanal dairy products are a gold mine in terms of diversity of strains and could be potentially interesting to dairy companies for the formulation of functional starter cultures in the production of innovative foods.     

Effect of Low-pressure Cold Plasma (LPCP) on the Wettability and the Inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> on Fresh-Cut Apple (<Emphasis Type="Italic">Granny Smith</Emphasis>) Skin

The aim of this study was to investigate the effect of low-pressure cold plasma (LPCP) on the inactivation of Escherichia coli and Listeria innocua on fresh-cut apple skin and its influence on wettability. Cold plasma treatments have shown to be effective to decontaminate foods, but their effect on the wettability has not been well studied. Surface-inoculated apple samples were treated with argon (Ar), nitrogen (N2), oxygen (O₂), and argon-oxygen (Ar-O₂) cold plasma using a commercial LPCP unit. Three different models were used to fit bacterial survival curves after the LPCP treatments. Changes in surface wettability were also determined by measuring the contact angle. The LPCP treatments using Ar, O₂, or Ar-O₂ mixture for 20 min were the most effective to inactivate E. coli with O₂, while the LPCP treatment with N2 for 20 min reduced L. innocua the most for (p?<?0.05). The highest increase in surface wettability was observed in samples treated for 20 min with O₂ and Ar-O₂. Different LPCP treatments have not great effectivity on the inactivation of E. coli and L. innocua on fresh-cut apple surface, but the all treatments changed the surface wettability of apples, making it more hydrophilic. This can be considered as a negative effect of the LPCP treatment because it can facilitate the adhesion and proliferation of re-contaminating microorganisms. More research should be undertaken to explore the use of other gases and complex surfaces.     

Effect of Amino Acids on Microwave Dielectric Properties of Minced Antarctic Krill (<Emphasis Type="Italic">Euphausia superba</Emphasis>)

The dielectric properties of ten solid amino acids, namely, glycine, proline, cysteine, glutamine, lysine, methionine, histidine, arginine, tyrosine, and tryptophan, and the effects of amino acid concentrations (0, 1, 3, and 5%) on the dielectric properties of minced Antarctic krill (Euphausia superba) were studied within a frequency range of 300–3000 MHz at 20.0 ± 0.5 °C. The dielectric constants and dielectric loss factor of amino acids were 2.0 ± 0.4 and 0.2 ± 0.3, respectively (moisture contents, 0.05–0.33%), except those of lysine (5.5 ± 0.5 and 1.5 ± 1.0, respectively) (moisture content, 4.5%). The dielectric properties of non-polar, neutral-polar, alkaline amino acids added to Antarctic krill were affected by the frequency and moisture contents, molecular weight, and isoelectric points (pI) of amino acids. As the molecular weight and pI values increased, the critical frequencies of aliphatic amino acid added to minced Antarctic krill decreased, whereas heterocyclic amino acid added to minced Antarctic krill increased. Quantitative relationships between parameters that define both the real and the imaginary parts of the Debye relations and the weight fraction of amino acid contents in Antarctic krill were determined. Moreover, glycine (at 5%) or lysine (at 5%) significantly influenced the dielectric constants and dielectric loss factor of minced Antarctic krill (p < 0.05) but did not have significant effects on temperature history and distribution (p < 0.05) during microwave heating at 2450 MHz.     

Development and characterization of reconstituted hydrogel from <Emphasis Type="Italic">Aloe vera</Emphasis> (<Emphasis Type="Italic">Aloe barbadensis</Emphasis> Miller) powder

Structural and rheological characterization of reconstituted hydrogels developed from A. vera non-fibrous alcohol insoluble residue (NFAIR) powder using different methods [viz., shaking (S), heating-shaking (HS), and heating (H)] and concentrations (viz., 0.2–1.6 %, w/v) was carried out. Functional group distribution by FTIR spectroscopy and Congo red (CR) method revealed the presence of acetylated acemannan in A. vera powder. Dynamic oscillation studies of A. vera (NFAIR) fluids at all concentrations of 0.2–1.6 %, w/v, showed gel strength in the order of H > HS > S method. However, in H method, increase in concentration from 0.2 to 1.6 %, w/v showed the conformational transition from semi-diluted solution to weak gel nature. Rheological models described the effect of heating temperatures (HT); 30–90 °C, and times (Ht); 15–60 min on viscoelastic behavior in reconstituted A. vera fluids. The reconstituted A. vera hydrogel prepared with a concentration of 1.6 %, w/v using 50 °C (HT) and 30 min (Ht) condition showed a good agreement with the Power law (storage modulus, G′) and Weak gel model (complex modulus, G*) fitted data (R² > 0.94) resulting higher viscoelastic moduli intercepts; G′₀ (71.5 Pa s ^n′), G″₀ (33.5 Pa s ^n″), lower slopes; n′ (0.22), n″ (0.06), higher network strength (A _F , 121.3 Pa s^1/z ) and number of network (z, 5.3) values. The obtained results suggested that heating at 50 °C/30 min can develop aqueous weak gel networks of A. vera with enhanced gel strength which may be utilized as a novel gelling agent for wide variety of targeted applications in food and pharmaceutical sectors.     

Effect of PrA encoding gene-<Emphasis Type="Italic">PEP4</Emphasis> deletion in industrial <Emphasis Type="Italic">S</Emphasis>. <Emphasis Type="Italic">cerevisiae</Emphasis> WZ65 on key enzymes in relation to the glycolytic pathway

S. cerevisiae proteinase A (PrA) is a member of the aspartic proteinase superfamily. The roles of PrA in S. cerevisiae physiology and cell metabolism are controversial. The objective of the current study was to elucidate the effects of the absence of PrA on key enzymes with regard to the glycolytic flux in industrial S. cerevisiae. The observed activities of hexokinase (HK), phosphofructokinase (Pfk) and pyruvate kinase (PYKi) in PrA-modified S. cerevisiae strains (SC2 and SC3) were lower than that of the wild-type strain (p < 0.01). Compared to other strains, SC3 revealed the lowest activity of three key glycolytic enzymes. Current results imply that PrA in industrial S. cerevisiae may control the glycolytic enzymes expression (HK, Pfk and PYKi) in the direct or indirect manner and thus lead to the delay of cell metabolism. The observed intracellular ATP levels between the wild-type strain and PrA-modified strains (SC1 and SC2) were significantly different (p < 0.01). The pronounced differences of key glycolytic enzymes between the wild-type and PEP4-modified strains were as well characterized by SDS–PAGE. The intracellular protein concentration in the presence of PrA is higher than those of the PrA-modified strains. As a result, the interaction mode of PrA and the glycolytic enzymes was postulated in this work. The findings herein suggest that the glycolytic flux direction and rate may be regulated by vacuolar PrA in industrial S. cerevisiae. The present data obtained provide insights into understanding the roles of PrA in industrial S. cerevisiae.