Fungus Geotrichum Candidum Research Articles

Fludioxonil does not antagonize propiconazole activity against Geotrichum candidum.

Fludioxonil and propiconazole are frequently used fungicides for managing fungal diseases in specialty crops. They are often used in mixture to manage brown rot caused by Monilinia fructicola and sour rot caused by the yeast-like fungus Geotrichum candidum,respectively, on stone fruits. Research indicated, however, that fludioxonil can increase the expression of the ATP-binding cassette transporters CDR1 and CDR2 in the yeast Candida albicans, leading to increased resistance to DMI fungicides. In this study, mycelial growth assays and detached fruit assays were conducted with G. candidum isolates of varying sensitivity to propiconazole to investigate potential antagonism between the two active ingredients. Isolates sensitive, reduced-sensitive, and resistant to propiconazole were able to grow with or without 20 µg/ml fludioxonil on PDA containing up to 0, 3, and 30 μg/ml propiconazole and on water agar containing up to 7.5, 7.5, and 30 μg/ml propiconazole, respectively, and no antagonistic interactions were detected. Propiconazole controlled disease with or without 20 µg/ml fludioxonil equally on detached cherry fruit caused by isolates with different DMI sensitivity phenotypes. Our results indicated no detectable antagonism between fludioxonil and propiconazole that would interfere with the mixture's ability to manage sour rot of cherry.

Addressing the Concern of Orange-Yellow Fungus Growth on Palm Kernel Cake: Safeguarding Dairy Cattle Diets for Mycotoxin-Producing Fungi.

Palm kernel cake (PKC), a byproduct of palm oil extraction, serves an important role in Ecuador's animal feed industry. The emergence of yellow-orange fungal growth in PKC on some cattle farms in Ecuador sparked concerns within the cattle industry regarding a potential mycotoxin-producing fungus on this substrate. Due to the limited availability of analytical chemistry techniques in Ecuador for mycotoxin detection, we chose to isolate and identify the fungus to determine its association with mycotoxin-producing genera. Through molecular identification via ITS region sequencing, we identified the yellow-orange fungus as the yeast Candida ethanolica. Furthermore, we isolated two other fungi-the yeast Pichia kudriavzevii, and the fungus Geotrichum candidum. Molecular identification confirmed that all three species are not classified as mycotoxin-producing fungi but in contrast, the literature indicates that all three have demonstrated antifungal activity against Aspergillus and Penicillium species, genera associated with mycotoxin production. This suggests their potential use in biocontrol to counter the colonization of harmful fungi. We discuss preventive measures against the fungal invasion of PKC and emphasize the importance of promptly identifying fungi on this substrate. Rapid recognition of mycotoxin-producing and pathogenic genera holds the promise of mitigating cattle intoxication and the dissemination of mycotoxins throughout the food chain.

Assessment and cost analysis of mycoremediated textile effluent for irrigation

AbstractIn the current study, mycoremediated textile effluents were used for irrigation in a cultivation system to promote resource recycling and cost‐effective spinach production. The raw textile effluents from two different textile mills were decolorized by 93.2 and 94.5%, respectively, as well as mineralized by the immobilized novel fungus Geotrichum candidum. Subsequently, both the treated effluents, treated textile effluent 1 (TTE1) and treated textile effluent 2 (TTE2), were used to cultivate spinach. The results indicated that irrigation with TTE1 and TTE2 resulted in ~9% higher spinach yields compared with the control. Moreover, the water use efficiency with the TTE1 and TTE2 treatments was ~1.2 times better than that with the control. The need for fertilizers in the case of spinach cultivation with treated effluents was compensated by the textile effluents as a result of the breakdown and mineralization of the effluent by immobilized G. candidum. Notably, the energy content, dietary fiber and protein content of spinach irrigated with mycoremediated textile effluent were substantially elevated, with values of 34 kcal/100 g−1, 3.4 g 100 g−1 and 6.92 g 100 g−1, respectively. The preliminary cost analysis indicated that the costs of the whole process (from treatment to irrigation) were approximately US$0.35‐0.38 m−3 (INR 28.22–30.45 m−3) of textile effluent. The study concluded that the mycoremediated textile effluent could be efficiently applied for agronomic usages.

High Inhibition for a CoII Tetrazole Bi‐pyrazole Dinuclear Complex against Fusarium Oxysporum f. sp. Albedinis

AbstractNew coordination compounds made of two novel tetrazole and C,N‐bipyrazole ligands, 2‐(3,5,5′‐trimethyl‐1′H‐[1,3′‐bipyrazol]‐1′‐yl)acetonitrile (L1), and 1′‐((1H‐tetrazol‐5‐yl)methyl)‐3,5,5′‐trimethyl‐1′H‐1,3′‐bipyrazole (HL2), were prepared and fully characterized by spectroscopic techniques. Their crystal structures were identified by single‐crystal X‐ray diffraction revealing mononuclear complexes: [Ni(L1)3](ClO4)2 (1), [Cd(L1)2Cl2] (2), [Cu(HL2)(L2)]ClO4 (3), [Cu(L2)2] (4) and a dinuclear complex [Co2(HL2)(L2)Cl3] (5) comprising CoII ions in octahedral and tetrahedral surrounding within the same unit. Noticeably, 3 and 4 show different architectural structures due to ligand deprotonation as well as the effect of the counter anion. All compounds demonstrated antimicrobial activity against Gram (+) bacteria Listeria innocua and Staphylococcus aureus, as well as Gram (−) bacteria such as Escherichia coli and Pseudomonas aeruginosa, and antifungal activity against pathogenic fungi Geotrichum candidum, Aspergillus niger, and Penicillium crustosum. Interestingly, a high inhibition activity of 97 % was reached for 5 with a low concentration of only 81.1 μmol/L against Fusarium oxysporum f. sp. Albedinis, which is commonly damaging palm trees crops.

Progressive Vertical and Horizontal Phytocompound Changes during Agarwood Formation in Aquilaria sinensis after Geotrichum candidum Injection.

(1) Background: Agarwood is an aromatic resin produced by the host tree through an immunological response against biotic and abiotic stress. The aim was, first, to use the fungus Geotrichum candidum to stimulate compound changes in Aquilaria sinensis horizontally (color formation) and vertically (cutting layers) after injection with it. (2) Methods: Horizontal and vertical sections were collected and separated five months after injection with the fungal broth. Two grams of dry powder was mixed with 20 mL methanol for 3 h at room temperature, and the solution was vibrated in an ultrasonic cleaner bath at 40 °C for 1 h. After vacuum drying, a concentration of 10 mg/mL of the tested samples in methanol was prepared for reversed-phase high-performance liquid chromatography (RP-HPLC), gas chromatography/mass spectrometry (GC/MS), and thin-layer chromatography (TLC) analysis. (3) Results: The horizontal changes in the compounds and their concentrations were associated with color. Compared to the normal (N) group, G. candidum injection stimulated more compounds at RT 27-42 in the white (W) group, brown (BR) group, and black (B) group. Furthermore, a significant increase in fatty acids was observed in the W group, implying an early plant response after G. candidum injection. In the BR group, the compounds were more similar to commercial agarwood (Out group). In the B group, alkaloids were the main compounds. Vertical changes in the main compounds were not observed, although the compound level varied. A TLC analysis determined the main compounds in the BR group at 254 nm and in the B group at 365 nm. Higher fatty acid levels were found in L6 and L5 and were correlated with higher terpenoid and sesquiterpene levels, suggesting that these compounds were possibly the first stage of agarwood formation. A GC/MS analysis demonstrated that the main compound groups were almost identical to the BR parts. (4) Conclusions: The injection of G. candidum led A. sinensis to synthesize different phytochemicals horizontally, not vertically, in the BR group.

Molecular Characterization of Two Totiviruses from the Commensal Yeast Geotrichum candidum.

Mycoviruses can infect many of the major taxa of fungi including yeasts. Mycoviruses in the yeast fungus Geotrichum candidum are not well studied with only three G. candidum-associated viral species characterized to date, all of which belong to the Totiviridae genus Totivirus. In this study, we report the molecular characteristics of another two totiviruses co-infecting isolate Gc6 of G. candidum. The two totiviruses were tentatively named Geotrichum candidum totivirus 2 isolate Gc6 (GcTV2-Gc6) and Geotrichum candidum totivirus 4 isolate Gc6 (GcTV4-Gc6). Both viruses have the typical genome organization of totiviruses comprising two ORFs encoding capsid protein (CP) and RNA-dependent RNA polymerase (RdRp) at the N and C termini, respectively. The genomes of GcTV2-Gc6 and GcTV4-Gc6 are 4592 and 4530 bp long, respectively. Both viruses contain the-frameshifting elements and their proteins could be expressed as a single fusion protein. GcTV2-Gc6 is closely related to a totivirus isolated from the same host whereas GcTV4-Gc6 is related to insect-associated totiviruses. The phylogenetic analysis indicated that GcTV2-Gc6 and GcTV4-Gc6 belong to two different sister clades, I-A and I-B, respectively. It is interesting that all viruses identified from G. candidum belong to the genus Totivirus; however, this might be due to the lack of research reporting the characterization of mycoviruses from this fungal host. It is possible that the RNA interference (RNAi) mechanism cannot actively suppress totivirus accumulation in G. candidum Gc6.

Domestication of different varieties in the cheese-making fungus Geotrichum candidum

Domestication of different varieties in the cheese-making fungus Geotrichum candidum

Impact of Dragon Fruit Waste in Microbial Fuel Cells to Generate Friendly Electric Energy

Pollution generated by the misuse of large amounts of fruit and vegetable waste has become a major environmental and social problem for developing countries due to the absence of specialized collection centers for this type of waste. This research aims to generate electricity in an eco-friendly way using red dragon fruit (pitahaya) waste as the fuel in single-chamber microbial fuel cells on a laboratory scale using zinc and copper electrodes. It was possible to generate voltage and current peaks of 0.46 ± 0.03 V and 2.86 ± 0.07 mA, respectively, with an optimum operating pH of 4.22 ± 0.09 and an electrical conductivity of 175.86 ± 4.72 mS/cm at 8 °Brix until the tenth day of monitoring. An internal resistance of 75.58 ± 5.89 Ω was also calculated with a maximum power density of 304.33 ± 16.51 mW/cm2 at a current density of 5.06 A/cm2, while the FTIR spectra showed a decrease in the initial compounds and endings, especially at the 3331 cm−1 peaks of the O–H bonds. Finally, the yeast-like fungus Geotrichum candidum was molecularly identified (99.59%). This research will provide great opportunities for the generation of renewable energy using biomass as fuel through electronic devices with great potential to generate electricity.

Uncovering the Biotechnological Importance of Geotrichum candidum

Fungi make a fundamental contribution to several biotechnological processes, including brewing, winemaking, and the production of enzymes, organic acids, alcohols, antibiotics, and pharmaceuticals. The present review explores the biotechnological importance of the filamentous yeast-like fungus Geotrichum candidum, a ubiquitous species known for its use as a starter in the dairy industry. To uncover G. candidum's biotechnological role, we performed a search for related work through the scientific indexing internet services, Web of Science and Google Scholar. The following query was used: Geotrichum candidum, producing about 6500 scientific papers from 2017 to 2022. From these, approximately 150 that were associated with industrial applications of G. candidum were selected. Our analysis revealed that apart from its role as a starter in the dairy and brewing industries, this species has been administered as a probiotic nutritional supplement in fish, indicating improvements in developmental and immunological parameters. Strains of this species produce a plethora of biotechnologically important enzymes, including cellulases, β-glucanases, xylanases, lipases, proteases, and α-amylases. Moreover, strains that produce antimicrobial compounds and that are capable of bioremediation were identified. The findings of the present review demonstrate the importance of G. candidum for agrifood- and bio-industries and provide further insights into its potential future biotechnological roles.

Sanitary and microbiological significance and features of identification of Geotrichum candidum

In the presented work we have studied the sanitary-microbiological, veterinary and medical significance of the mycelial fungus Geotrichum candidum. Its morphological, cultural and tinctorial properties were determined, and the method of identifying mold fungi in the scotch preparation was optimized. Pure Geotrichum candidum culture was obtained and studied during the study of the fermented milk product matsoni, the number of hyphae of mold fungus was determined according to Howard, the quality and degree of spoilage of the food product of liquid consistency were assessed.

Identification of novel totiviruses from the ascomycetous fungus Geotrichum candidum.

Mycoviruses are widely distributed across the kingdom Fungi, including ascomycetous yeast strains of the class Saccharomycetes. Geotrichum candidum is an important fungal pathogen belonging to Saccharomycetes and has a diverse host range. Here, we report the characterization of four new classical totiviruses from two distinct Geotrichum candidum strains from Pakistan. The four identified viruses were tentatively named "Geotrichum candidum totivirus 1, 2, 3a, and 3b" (GcTV1-3b). The complete dsRNA genomes of the identified totiviruses are 4621, 4592, 4576, and 4576 bp in length, respectively. All totivirus genomes have two open reading frames, encoding a capsid protein (CP) and an RNA-dependent RNA polymerase (RdRP), respectively. The downstream RdRP domain is assumed to be expressed as a CP-RdRP fusion product via -1 frameshifting mediated by a heptameric slippery site. Sequence comparisons and phylogenetic analysis showed that each of the discovered viruses belongs to a new species of the genus Totivirus in the family Totiviridae, with GcTV1 and GcTV3 (a and b strains) clustering in one subgroup and GcTV2 in another subgroup.

How do the Growth and Metabolic Activity of Aquatic fungi Geotrichum Candidum and Aspergillus Niger Respond to Nanoplastics?

In this study, exposure experiments were conducted to assess the effects of polystyrene nanoparticles (PS) and amine-modified polystyrene nanoparticles (APS) at environmental concentrations (1, 10, and 100µg L- 1) on two fungal species (Geotrichum candidum and Aspergillus niger), isolated from leaf litter in streams, concerning their growth and metabolic activity. Results showed that PS at 1 and 10µg L- 1 have hormesis effects on G. candidum growth. Compared with G. candidum, A. niger had higher sensitivity to nanoplastic exposure. Besides, the peroxidase and cellobiohydrolase activities of A. niger were significantly inhibited by nanoplastics (except 1µg L- 1 PS), which would weaken its metabolic activity in carbon cycling. These results provided a new thought on how the growth and functions of aquatic fungi cope with the stress induced by nanoplastics. Overall, the study provided evidence for the different responses of aquatic fungi to nanoplastics in streams.

Dendrobaena veneta avoids ethyl pentanoate and ethyl hexanoate, two compounds produced by the soil fungus Geotrichum candidum.

Earthworms shape the biological and physicochemical qualities of the soil they choose to reside in, but our understanding of the specific chemicals that attract or repel a particular species of earthworm remains incomplete. Current research indicates that some species feed on and are attracted to fungi, such as Geotrichum candidum. In the present study, as part of our continuing effort to characterize mechanisms of earthworm chemosensation, we tested whether ethyl hexanoate and ethyl pentanoate, two compounds produced by G. candidum, are appetitive to the European nightcrawler (Dendrobaena veneta). In a soil T-maze, both of these compounds significantly repelled individual earthworms in a dosage-dependent manner, this result ran counter to our initial hypothesis. D. veneta also avoided ethyl hexanoate and ethyl pentanoate in an assay we specifically developed to test an earthworms aversion to chemical stimuli in soil. In both of these assays, ethyl hexanoate was aversive at lower concentrations than ethyl pentanoate. These findings further clarify our understanding of the chemical cues that trigger the decision of D. veneta to select a particular soil-environment, and emphasize that different earthworm species may react very differently to commonly encountered chemical stimuli.

Immobilized fungal consortium‐GR: a novel textile effluent treatment alternative for removal of azo dyes and other pollutants

AbstractBACKGROUNDCleaning up the environment by the elimination of harmful contaminants from textile effluents (TE) is a critical and challenging crisis that requires long‐term appropriate solutions. In this work, a fungal consortium‐GR containing two fungi Geotrichum candidum F4 and Rhodotorula glutinis F7, in the ratio 1:1, is demonstrated to treat the organic contaminants in TEs.RESULTSThe immobilized consortium‐GR in coconut fibre is 24% more efficient at treating TE than free consortium‐GR, showing a maximum decolorization of 88.4% in 6 h immersion, at optimal conditions of pH 5.5, temperature 35 °C, inoculum size 5% and fibre quantity 2.5 g L−1. Biodegradation was confirmed by using UV–visible spectrophotometry, high‐performance liquid chromatography and Fourier transform infrared spectroscopy, and the degradation pathway of methyl orange was confirmed by gas chromatography–mass spectroscopy study. The novel immobilized consortium‐GR in the bioreactor was assessed by the reduction of toxic parameters in 6 h: colour by 70%, total dissolved solids by 62%, total suspended solids by 81%, biological oxygen demand by 51%, chemical oxygen demand by 58%, chloride by 78%, ammonia by 64%, sulfate by 55%, lead by 12.5% and chromium by 75%. During this process of bioconversion, maximum laccase activity of 3.25 U L−1 was observed in 6 h by immobilized consortium‐GR. Phytotoxicity and genotoxicity studies revealed the detoxification of TE.CONCLUSIONThrough the continuous reuse of treated water in subsequent dyeings, the proposed TE treatment process represented a 92.86% reduction in water use and a total cost reduction of 22.47%.This study is a step towards upcoming onsite, field and short‐term applications of consortium‐GR for the management of TE. © 2021 Society of Chemical Industry (SCI).

Current Status, Economic importance and Management of Dodders (Cuscuta Spp) of Important Crops

Dodders ( Cuscuta spp.) are one of the worst weeds that causes economic damage to economically valued crops. Cuscuta campestris , C. kilimanjari, C. reflexa, C. australis , C. suaveolens , C. hyalina , C. cassytoides , C. epilinum and C. planiflora are the most widespread Cuscuta species globally. Most of them are originated in North America and later distributed to many countries in the world. Coffea spp., Camellia sinensis, Glycine max, Allium cepa and Phaseolus spp. are the major hosts of the weed . Dodders are holoparasites, which lack chlorophyll and derive all their resources from their host and impose serious damage to their hosts. Cultural practices such as planting non-host cereal crops by removing host plants; regular monitoring and removal of new dodder weeds and mixed cropping of host crop with non-host crops is effective in managing the weed. Besides, biological control by use of parasitic fungi ( Alternaria destruens , Fusarium tricinctum, Alternaria alternata , Geotrichum candidum and Alternaria spp and Colletotrichum gloeosporioides ) play a role in managing the weed. In addition, chemical control with pre-emergence herbicides (pronamide, trifluralin and pendimethalin) and post-emergence herbicides (pelargonic acid, imazamox, imazethapyr and paraquat) are effective to control dodders in many crops in extreme cases. Integrated use of herbicidal and mechanical control measures is effective in control, economically safe, socially acceptable and environmentally friendly than a single control measure. Moreover, host specific integrated Cuscuta Spp . management should be designed. Keywords: Cuscuta campestris, Hautoria, Holoparasite, Parasitic weeds, Stem parasite DOI: 10.7176/ALST/87-04 Publication date: April 30 th 2021

Growth dynamics of lactic acid bacteria and dairy microscopic fungus Geotrichum candidum during their co-cultivation in milk.

Production of high-quality and microbiologically safe fermented dairy products requires controlled growth and microbial interactions between lactic acid bacteria and microscopic fungi. For this purpose, detailed knowledge of their growth characteristics is needed. Therefore, the objective of this study was to analyse the growth dynamics of lactic acid bacteria of commercial DVS® FRESCO® 1000NG culture and dairy isolate of microscopic fungus Geotrichum candidum during their co-cultivation in milk. The growth dynamics of microorganisms was studied in dependence on their initial counts at 12, 15, 18, 21 and 30 °C. Growth parameters were calculated by two primary predictive models, model of Baranyi and Roberts and Huang's model. Both models showed good ability to describe the growth dynamics of studied microorganisms, as it was confirmed by low values of RMSE index. Both microbial cultures, Fresco culture and Geotrichum candidum, showed good growth ability in milk since they reached the average maximum density of 9.50 ± 0.13 log CFU/mL and 5.85 ± 0.69 log CFU/mL (n = 45), in order. Maximum density of studied microorganisms was not affected by their initial counts or incubation temperature. On the other hand, effect of mutual ratio of microbial initial counts and increasing temperature had a significant impact on growth dynamics.

Modification of secondary products of processing triticale into starch with a new strain of the fungus Geotrichium candidum

The aim of this work was to study the possibility of using a new strain of the fungus Geotrichum candidum for the bioconversion of serum remaining after the isolation of starch and proteins from triticale grain. The fungus strain Geotrichum candidum 977 was isolated from the steep waters of triticale grains formed during the production of starch and identified on the basis of analysis of the sequence of ribosomal genes. The strain was characterized by large cells efficient separation of biomass from the culture liquid and high growth rate. On protein-free whey, which remains after isolating proteins from steeping waters the fungus assimilated glucose, maltotriose, fructose and did not assimilate maltose. At a pH of 5.0 the growth of the fungus was not observed at a pH of 5.5 to 6.5 it was weak at a pH of 7.5 to 8.5, the productivity of the fungus increased 1.8 times and amounted to 3.00-3,15 g / 100 cm3. During growth the strain alkalized the medium from pH 5.5 to pH 8.5. The microbial-plant concentrate contained 33.3 ± 2.1% protein and 19 amino acids with a predominance of alanine, aspartic, glutamic acids, lysine, threonine and leucine. The score of essential amino acids exceeded 100% with the exception of sulfurcontaining ones (64 -72%). Thus, the possibility of using a new strain of the fungus G. Candidum 977 for the utilization of wastewater generated during the production of starch and proteins from triticale grains has been shown to obtain fodder protein concentrates.

BIOTRANSFORMATION OF COMPONENTS OF PEA GRAIN IN FOOD AND FODDER PROTEIN CONCENTRATES

The data were obtained on the development of biotechnology of food protein concentrate from pea grains and bioconversion of cereal serum with Saccharomyces cerevisiae 121 yeast and a new strain of the fungus Geotrichum candidum 977 with the formation of a feed microbial-plant preparation.

Зерновые композиты с комплементарным аминокислотным составом для пищевых и кормовых целей

The present paper features processes of serum biotransformation. The serum was obtained from triticale extract and pea flour after protein concentrates of increased biological value had been extracted. The research objective was to obtain microbial and vegetable feed concentrates by using a composition of Saccharomyces cerevisiae121 yeast and the yeast-like fungus Geotrichumcandidum 977. The mass fraction of protein in the two-component composites was 75–80% of the dry matter. The score of the first and the second limiting amino acids (lysine and threonine) equaled 103–113%, and that of the third acid (sulfur- containing) was 71–72%. The chemical composition of the composites corresponded to the ‘Concentrates’ group; the values of their functional and technological properties were typical of concentrates from other types of grain crops. The study revealed some cultures that are able to actively develop in serum, which is a secondary product of processing the extract after protein isolation. A symbiotic ferment was prepared from the fungus Geotrichumcandidum 977 and the yeast Saccharomyces cerevisiae 121, which ensures the growth of biomass in a carbohydrate- and nitrogen-containing medium. Proteins were isolated under the action of amylase, glucoamylase, cellulose, and xylanase. The amount of high-molecular compounds (dextrins) and trioses (raffinose) released from the interaction with protein and non-starch polysaccharides decreased 2–4 times in the solution. The amount of glucose, disaccharides, xylose, and galactose increased 2–10 times, compared with the original extracts. The serum remaining after the removal of the main mass of the protein was enriched with low molecular weight mono- and oligosaccharides, which positively affected the growth of microorganisms. The mass fraction of proteins in the microbial-vegetable composite obtained from the extract with the triticale proteins and pea flour ratio of 1:5 was 15% higher than at the ratio of 1:3. Microbial and vegetable concentrates with a mass fraction of protein of 55.8–75.1% of dry matter can be used in fodder production as a protein-carbohydrate additive. Protein composites made of protein triticale and peas with a complementary amino acid composition can improve the biological value and performance of food products.

Immobilized and Free Cells of Geotrichum candidum for Asymmetric Reduction of Ketones: Stability and Recyclability.

Marine-derived fungus Geotrichum candidum AS 2.361 was previously reported by our group as an active strain for the enantioselective reduction of ketones. Although some other Geotrichum strains were also found from the terrestrial sources, information on their stability and reusability is scarce. Herein, the stabilities—in terms of pH tolerance, thermostability, and storage stability, and reusability—of G. candidum AS 2.361 were described for the asymmetric reduction of a series of aromatic ketones. Two differently immobilized cells (agar immobilization and calcium alginate immobilization) as well as free cells were prepared. For three substrates (1-(3-bromophenyl) ethan-1-one (1b), 1-(2-chlorophenyl) ethan-1-one (1d), and acetophenone (1g)) immobilized cells on agar showed a great improvement in the bioreduction activities compared to the free cells, increasing yields up to 97% with ee values of 99%. Cells immobilized on agar/calcium alginate could maintain more than 90% of the original activities within the assayed pH ranges of 3.5–11, while free cells were highly sensitive to alkaline and acidic conditions. Concerning thermostability, immobilized cells on agar kept 99% of their original activities after incubation at 60 °C for 1 h, while almost no activity was detected for the free cells under the same condition. Immobilized cells were stable at 4 °C for 80 days without any activity loss, while free cells started to decrease the activity after storage at 4 °C for six days. The immobilized cells retained almost 99% activity after four reuse cycles, while free cells lost almost all the activities at on the third cycle.