Extraction and characterization of a novel <Emphasis Type="Italic">Terminalia</Emphasis> pectin

This study reports the investigation into pectin present in Terminalia ferdinandiana—a native Australian fruit utilised in dietary supplement industry. Citric acid extraction was carried out to extract pectin from two commercially available T. ferdinandiana products—frozen puree and freeze-dried puree powder. The yields of the extracted pectin were measured at various pHs (2.0, 3.0 and 4.0) and times (30, 60 and 120 min) at 75 °C. Pectin yield ranged between 4.8 and 21%. Freeze-dried powder had a higher pectin yield compared to puree. Extraction at pH 3 for 120 min resulted in the highest yield from both puree (15%) and powder (21%). T. ferdinandiana pectins were found to have low methoxyl content with degree of esterification of 35.07 and 34.74% for puree and powder, respectively. Fourier transform infrared spectroscopy confirmed that the extracts were pectin.     

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.     

Effects of Vacuum Impregnation with Calcium Ascorbate and Disodium Stannous Citrate on Chinese Red Bayberry

This study aimed to improve the shelf life of Chinese red bayberries using vacuum impregnation. Vacuum pressure of 5 kPa for 15 min, atmospheric pressure for 10 min, an impregnation temperature of 20 °C, alone or in combination with isotonic sucrose solution, 1% food-grade disodium stannous citrate (DSC) and 2% food-grade calcium ascorbate were used for vacuum impregnation. Quality parameters, including firmness, weight loss, decay rate, microbial counts and polyphenol oxidase (PPO) and peroxidase (POD) activities, of red bayberries were studied at 2 °C for 10 days. The monosaccharide components, chemical structures and nanostructure properties of chelate-soluble pectin (CSP) were further studied using high-performance liquid chromatography, Fourier transform infrared spectroscopy and atomic force microscopy (AFM). The results indicated that vacuum impregnation with calcium ascorbate alone or calcium ascorbate combined with DSC showed significant effects on inhibiting the decrease of firmness (4–10 days), the increase of weight loss (2–10 days), decay rate (4–10 days) and microbial growth (2–10 days). In addition, vacuum impregnation with single calcium ascorbate or DSC or their combination significantly inhibited the increase of colour difference from day 6 to day 10 during storage, which was better than atmospheric impregnation. Furthermore, vacuum impregnation with DSC and calcium ascorbate had the best effect on sensory attributes. The nanostructure analysis by AFM showed CSP of large width and length when calcium ascorbate was impregnated, suggesting that vacuum impregnated with 2% calcium ascorbate inhibited the degradation and dissociation of CSP, although these fruits showed more branching of rhamnogalacturonan and a small change in chemical structure.     

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.     

Optimization of Trehalose Production by <Emphasis Type="Italic">Rhodotorula dairenensis</Emphasis> Following a Sequential Strategy of Experimental Design

Initially, a technical screening was applied to select yeasts with a potential for trehalose production using thermal stress for the induction of synthesis. The effects of the concentrations of sugar cane molasses, corn steep liquor and a commercial yeast extract Prodex Lac SD®, and of pH and temperature on the biomass were studied using a fractional design followed by a central composite rotatable design (CCRD). The optimum values were 50 g/l for the molasses and corn steep liquor concentrations, initial pH of 5.5, and temperature of 30 °C, with no yeast extract. Under these conditions, the production of trehalose was studied using a CCRD to optimize the temperature (33 to 47 °C) and exposition time (60 to 120 min) of the cultures to the thermal stress. The maximum intracellular trehalose content reached was 20.5% (g trehalose/100 g dry cell) at temperatures of 35–40 °C and with 100–120 min of exposition time to the thermal stress.     

Effect of vacuum impregnation on the phenolic content of Granny Smith and Stark Delicious frozen apple <Emphasis Type="Italic">cvv</Emphasis>

ABTS•+ test, o-diphenols (spectrophotometric) and HPLC-DAD phenolic content in vacuum impregnated apple slices from Stark Delicious and Granny Smith cvv. were studied. Vacuum impregnation (VI) was carried out in a pilot plant using an aqueous solution containing 37.9% dextrose, 15.2% sucrose, 1.0% ascorbic acid, 0.25% calcium chloride and 0.25% sodium chloride. The impregnation time was 30 min and the vacuum pressure was 100 mbar; the temperature was 25 °C during all processing steps and the solution/fruit ratio was 11:1. At the end of VI, samples were left 5 min in syrup. They were then rinsed with water, drained in a vibrating screen and quickly frozen. A significant decrease (p < 0.05) of the o-diphenol content was seen (17.84 and 12.32% of the initial content in Stark and Granny varieties, respectively). The same trend was confirmed by HPLC-DAD where reductions in total phenols were 21.57 and 26.86% in Stark and Granny, respectively. Individual phenolic compounds showed different rates of depletion, although in some cases there was no reduction. The ABTS•+ test was strongly affected by the presence of ascorbic acid (AA), which was much higher in treated samples. Sensory evaluation showed higher values of hardness, crispness, juiciness and sourness in VI Granny Smith than VI Stark delicious, with a higher retention of texture parameters in the former. Finally, sweetness was slightly higher in VI Stark. An erratum to this article can be found at     

High Hydrostatic Pressure and Temperature Applied to Preserve the Antioxidant Compounds of Mango Pulp (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.)

High hydrostatic pressure (HHP) processes combined with moderate heating can be used to preserve foods while maintaining general quality. The effect of these conditions on the total phenolic (TP), vitamin C (L-ascorbic acid (AA)), carotenoids, and antioxidant activity (AOA) of mango purees was evaluated. Purees were processed at 400–550 MPa/34 and 59 °C at different holding times. Unprocessed puree had TP of 26.6 mg gallic acid/100 g, 21.1 mg L-ascorbic acid/100 g, AOA of 885 μmol trolox equivalents/100 g, and total carotenoids of 6.0 mg β-carotene/100 g. HHP treatments increased the phenolic concentrations up to 34% (550 MPa/59 °C/2 and 4 min) compared with the initial content, probably due to improvement of their extraction. AA content was reduced significantly (10–45%) after all HHP processes performed at 59 °C, while at 34 °C, they were diminished only after 8 and 16 min of treatment (13–26%). At 34 °C and lower times, AA concentration increased in average 18%. Total carotenoid retention in HHP-treated samples varies from 77 to 98%, being the higher the temperature the lower the retention observed. The concentration of most individual carotenoids remains unchanged, but violaxanthin content was reduced (21–26%) and 9-cis-violaxanthin was increased by about 10%. The AOA was also increased (up to 39%) at some processing conditions. A linear correlation between the TP and AOA was obtained. HHP at 550 MPa combined with moderate temperature (34 °C) at processing times up to 8 min is recommended for the maximum retention of the antioxidant compounds of mango puree.     

Comparative inactivation studies of <Emphasis Type="Italic">Aegle marmelos</Emphasis> (<Emphasis Type="Italic">bael</Emphasis>) peroxidase in crude extract of fruit by heat processing and ultrasonication treatment

Bael (Aegle marmelos) is considered as a holy fruit comprised of vast number of phytonutrients. Whole bael tree including all its parts has medicinal significance. Lack of awareness and seasonal nature makes its processing rather challenging. Conventional heat processing may lead to inactivation of quality hampering enzymes such as peroxidase, but at the cost of loss in essential phytonutrients. In the present work it was observed that thermal inactivation of bael peroxidase obeyed first order kinetics with enzyme activation energy of 7.7 kJ mol^?1. Complete inactivation of bael peroxidase was achieved within 11 min at 85?°C while ultrasound treatment attained in lesser time of 4 min at 64.07 W cm^?2 ultrasonic intensity. Loss of marmelosin a well-known phytonutrient in bael fruit was found to be 83.29?% by heat (11 min, 85?°C) and only 50.20?% by ultrasonication (4 min, 64.07 W cm^?2 ultrasonic intensity). Ultrasonication has potential to overcome harmful effects of heat processing with retention of phyto-constituents and hence has promising future in various food processing applications.     

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.     

Ultrasound-assisted extraction of bioactive compounds from <Emphasis Type="Italic">Malva sylvestris</Emphasis> leaves and its comparison with agitated bed extraction technique

The effects of ultrasound-assisted extraction (UAE) variables—namely extraction temperature (40–60 °C), ultrasonic power (50–150 W), and sonication time (40–60 min)—on the extractive value (EV) of bioactive phenolics from Malva sylvestris leaves were investigated and optimized using Response surface methodology. The effects of extraction solvents (ethanol, ethyl acetate, and n-hexane) on EV, free radical scavenging activity (FRSA), total phenolic content (TPC), and major bioactive phenolics were studied using agitated bed extraction (ABE), and the results were compared with the UAE findings. Under the optimal UAE conditions (48 °C, 110.00 W, and 48.77 min) the experimental EV was 279.89 ± 0.21 mg/g with 71.12 ± 0.15% DPPH_sc, 73.35 ± 0.11% ABTS_sc, and a TPC of 152.25 ± 0.14 mg GAE/g. Ethanolic ABE results in higher EV (320.16 ± 0.25 mg g^?1) compared to UAE, while the FRSA and TPC values were reduced. HPLC analysis revealed that the concentration of bioactive phenolics increased significantly (p < 0.05) under the optimal UAE conditions.     

Characterization and transformation of the <Emphasis Type="Italic">Opuntia ficus indica</Emphasis> fruits

Opuntia ficus indica fruits have been associated with health effects, due to their protective actions against oxidation. Nevertheless, few studies about processing of Opuntia fruits are available’; therefore, we studied the pulp characteristics and processing of a local variety, for producing beverage nectars. The pulp had an average pH of 5.64, 13.47 °Brix, with total sugars (106 g L^?1), K (1180 mg L^?1), 503.3 µg L^?1 of β carotene, 120 mg L^?1 of total phenolic compounds, 4.9 mg and 46.9 mg L^?1 respectively for betacyanins and betaxanthins and 243.4 mg L^?1 of vitamin C. The formulated nectars with 35% of pulp (N35) and 45% of pulp (N45) had respectively 14 and 15 °Brix. Minor components represent 1109 and 1112 mg L^?1 of K for N35 and N45 respectively, β carotene (318.6 µg and 362.8 µg L^?1), and vitamin C 227 and 231 mg L^?1. We confirmed the stability and acceptability of these beverages after a month of storage, after stability and panel tests. Therefore, we suggest that the pulp processing can be used as a new form of agro industrial utilization of this underutilized fruit.     

Optimization for encapsulation of green tea (<Emphasis Type="Italic">Camellia</Emphasis><Emphasis Type="Italic">sinensis</Emphasis> L.) extract by spray drying technology

Green tea has been credited with providing a wide variety of health benefits like higher total antioxidant, cancer prevention, and anti-irritant etc. Microencapsulation technology is used to prevent antioxidant loss during processing which involves entrapment of active material into carrier material. The experiments were designed using response surface methodology based on three level two factor (green tea extract, 10–25 % and drying inlet air temperature, 120, 130 and 140°C) central composite design. The feed emulsion was prepared with green tea extract as core material and 40 % maltodextrin concentration as wall material with the ratio of core to wall (1:2) and it was fed into the spray dryer at varied drying inlet air temperature to get encapsulated green tea extract powder. The quality characteristics were analysed such as moisture content, total antioxidant activity and total phenolic content and the process conditions were optimized using desirability function methodology. At optimum spray drying condition moisture content, total antioxidant activity and total phenolic content were found to be 3.1 % w.b., 72.91 % free radical scavenging activity by DPPH method, 57.81 mg/g of dry matter, expressed as gallic acid equivalent respectively with desirability value of 0.92.     

Bioactive properties of Kilka (<Emphasis Type="Italic">Clupeonella cultriventris caspi</Emphasis>) fish protein hydrolysates

Whole common Kilka fish was hydrolyzed separately using four commercial enzymes, Alcalase, Neutrase, Protamex at 50 °C and Pepsin at 37 °C for 30, 60 and 90 min. Degree of hydrolysis, angiotensin-I-converting enzyme (ACE) inhibitory activity and antimicrobial activity of each hydrolysate against Gram-negative (Escherichia coli, Salmonella enteritidis) and Gram-positive (Staphylococcus aureus, Listeria innocua) bacteria were studied. Results showed that the degree of hydrolysis for all enzymes was in the range of 2.63–3.36%. Electrophoresis profiles of the Kilka protein hydrolysates showed that most of produced peptides were in the range of 30 D but Alcalase and Neutrase had a better performance in the production of low molecular weight peptides in the range of 10 D. This led to increase the antimicrobial activity against the examined bacteria at the concentration of 200 µg/mL peptide solution. The Neutrase enzyme produced hydrolysate with the highest ACE inhibitory activity (53%?±?1.8 at 500 µg/mL). Antimicrobial activity of Kilka protein hydrolysates using Protamex and Pepsin was lower than the others due to lack of considerable amount of small peptides. The current research has demonstrated that the peptides derived from the enzymatic hydrolysis of Kilka fish protein in optimum conditions are capable of being converted to antimicrobial and antihypertensive agents to be used in functional foods.     

Infrared Heating as a Disinfestation Method Against <Emphasis Type="Italic">Sitophilus oryzae</Emphasis> and Its Effect on Textural and Cooking Properties of Milled Rice

Infrared (IR) heating method against rice weevils (Sitophilus oryzae) in an egg stage was investigated. A kinetic model was developed to describe insect mortality in a temperature range of 40–60 °C. Effects of IR heating temperature (50–60 °C) and exposure time (1–3 min) on insect mortality and quality attributes of the treated rice were evaluated. The optimized condition obtained by means of the response surface method was used to analyze rice quality before and after IR treatment with storage. The results showed that the 0.5th-order thermal death kinetic equation was the most suitable model, and the S. oryzae eggs had less heat tolerance than the adults and some other species. Mortality achieved 100 % after 2 min for all temperatures. Both IR heating parameters significantly affected the treated milled rice qualities. The minimal changes in rice quality before and after storage could be obtained using optimized temperature and exposure time of 53.6 °C and 1.2 min, respectively.     

Characterization of mannanase from <Emphasis Type="Italic">Bacillus</Emphasis> sp., a novel <Emphasis Type="Italic">Codium fragile</Emphasis> cell wall-degrading bacterium

Seaweeds are considered as a health food partly due to the polysaccharide composition of the cell wall. Because conventional extraction methods have low yields and lead to environmental pollution, enzymatic methods have been proposed. In this study, a new strain of Bacillus sp. was isolated from cattle feces that produced a mannanase, a polysaccharide-degrading enzyme active against the green seaweed Codium fragile. The purified 39-kDa mannanase exhibited maximum activity at 55 °C and pH 6.0, and maintained its catalytic activity stably at temperatures up to 60 °C and at a broad pH range (5.0–11.0). Enzymatic activity was slightly enhanced by Cu²⁺ and Na⁺ but strongly inhibited by Fe²⁺, Ag⁺, and EDTA. The mannanase showed the highest specificity to the inexpensive substrates such as konjac powder and locust bean gum, and efficiently released various manno-oligosaccharides. This novel mannanase can thus be applicable in the food, feed, and pulp industries.     

Effect of Solids Concentration on the Physicochemical and Flow Properties of Cactus Pear Juices of Two Varieties (<Emphasis Type="Italic">Opuntia ficus-indica</Emphasis> and <Emphasis Type="Italic">Opuntia streptacantha</Emphasis>)

Juices from two varieties of cactus pear, a green (Opuntia ficus-indica) and a red (Opuntia streptacantha), were obtained and concentrated by evaporation. Both fruit varieties and their juices at different concentrations were characterized. Green cactus pears had significantly higher amount of pulp than red cactus pears; the peel of O. ficus-indica represented only 38 versus 52 % of the fruit for the O. streptacantha. Both varieties had no significant differences on moisture, density, pH, and titratable acidity, in contrary to soluble solids. Juice was concentrated under vacuum conditions to reach a final concentration of 42, 53–55, and 58–60 °Brix, respectively, and stored under refrigeration (10 °C) during 4 weeks. Physicochemical properties of the pears and juices were determined as fresh items (time zero) and every week for the concentrate juices through storage; similarly, flow parameters were measured at 10 and 25 °C. Concentrate density (1160–1283 kg/m³) was mainly affected by final soluble solids, while pH and acidity were affected differently depending on the variety. Concentrated juices at 42 °Brix were considered with Newtonian behavior with a viscosity of 2–22 mPa s, while those at higher concentrations were of pseudoplastic nature (n < 1.0 and K > 69 mPa sⁿ). Power Law model fitted better the flow behavior than Herschel-Bulkley model of concentrates of both varieties. Temperature, solid concentration, and/or storage time affected the consistency coefficient (K) and flow index (n) depending on the cactus pear variety. Overall, those concentrated juices from O. streptacantha were more stable and exhibited lower apparent viscosity.     

Ultrasound-assisted extraction and antioxidant activity of phenolic and flavonoid compounds and ascorbic acid from rugosa rose (<Emphasis Type="Italic">Rosa rugosa</Emphasis> Thunb.) fruit

The present study aimed to extract total phenolic compounds (TPC), total flavonoid compounds (TFC), and ascorbic acid (AA) from the fruit of rugosa rose (Rosa rugosa Thunb.) by ultrasound-assisted extraction (UAE), and to evaluate their antioxidant activities. UAE significantly increased the extract yield compared with that obtained using the conventional method. TPC, TFC, and AA were extracted, depending on the extraction conditions (temperature, time, and ethanol concentration), in the range of 50.73–96.69, 15.93–31.88, and 3.06–6.08 mg/g, respectively. TPC and TFC were effectively extracted at a relatively high temperature (50 °C) than AA was (30 °C). The solvent condition used to extract TPC, TFC, and AA was 50% ethanol. The UAE condition for the highest antioxidant activity was obtained 30 °C, 30 min, and 50% ethanol, which were the same condition for the highest AA extraction. Among the extracts, AA showed a strong correlation with antioxidant activity at p-value of 0.001.     

Effect of Chitosan Coatings Enriched with Lauroyl Arginate Ethyl and Montmorillonite on Microbial Growth and Quality Maintenance of Minimally Processed Table Grapes (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L. Kyoho) During Cold Storage

The effects of lauroyl arginate ethyl (LAE), chitosan (CTS), and CTS-containing montmorillonite and LAE coatings (CML) on the quality maintenance and shelf life of minimally processed grapes stored at 4 °C for 20 days were investigated. Compared with the control, LAE, and pure CTS coatings, the CML coatings effectively decreased grape weight loss, respiration, and decay incidence as well as microorganism counts of total aerobic bacteria, yeasts, molds, and lactic acid bacteria in grapes. The CML coatings also showed better effects on delaying the tissue softening and browning due to the decreased activities of polygalacturonase, pectin methyl esterase, and polyphenol oxidase. CML coatings did not significantly change the content of soluble solids and the titratable acidity or the pH value of grapes, compared with other samples. However, CML coatings ensured higher content of ascorbic acid and sensory quality in grapes. The results confirmed that CML coatings improve microbiological safety and preserve nutritional and sensory quality of minimally processed grapes.     

Studies on the storage stability of fermented red dragon fruit (<Emphasis Type="Italic">Hylocereus polyrhizus</Emphasis>) drink

The objective of this work was to study the effect of storage temperatures and duration on the stability of fermented red dragon fruit drink (FRDFD) on its betacyanins content, physicochemical and microbiological qualities (BPM) and determining sensory acceptability. Results showed that both storage temperatures and duration have a significant effect on betacyanins content and physicochemical properties of FRDFD. Aerobic mesophilic and yeast and mold counts were lower than 1?×?10³ CFU/mL for FRDFD stored at both temperatures. The loss of betanin (16.53–13.93 g/L) at 4 °C was 15.73% with no significant changes in physicochemical properties from week two onwards compared to 56.32% (16.53–7.22 g/L) of betanin loss at 25 °C. At week eight, FRDFD stored at 4 °C still contained 13.93 g/L betanin with a pH value of 3.46, suggested its potential as a functional drink which is sensory acceptable (mean score?>?80% using hedonic test) among consumers.     

Probe-Based Fluorescence Melting Curve Analysis for Differentiating <Emphasis Type="Italic">Larimichthys polyactis</Emphasis> and <Emphasis Type="Italic">Larimichthys crocea</Emphasis>

Larimichthys polyactis (redlip yellow croaker) and Larimichthys crocea (large yellow croaker) are commercially important fish species in East Asia with high differences in their market values. In Korea, consumers prefer L. polyactis to L. crocea, although it is difficult to distinguish them based on their morphological traits. The objective of this study was to develop an assay for differentiating L. polyactis and L. crocea using fluorescence melting curve analysis (FMCA) with a single locked nucleic acid (LNA) probe. Species-specific regions of the mitochondrial 16S rDNA were selected as LNA probes. The target sequences of L. polyactis and L. crocea had a 2-bp difference, and a single LNA probe was identified using melting temperature (Tm) shift. LNA probe was 100 % complementary to the target sequence of ten L. polyactis samples, giving a significantly higher Tm value (66 °C) than that of five L. crocea samples (42 °C). Overall, the developed LNA-based FMCA system had high efficiency, multiplexity, and simplicity, and could be effectively used for differentiating L. polyactis and L. crocea, and as a food analyzing method based on DNA sequence.