Fructosan form Paenibacillus kribbensis PS04 enhance disease resistance against Rhizoctonia solani and tobacco mosaic virus

BACKGROUND: Rice sheath blight (caused by Rhizoctonia solani) and tobacco mosaic virus are very important plant diseases, causing a huge loss in global crop production. Paenibacillus kribbensis PS04 is a broad-spectrum biocontrol agent, used for controlling these diseases. Previously, extracellular polysaccharides (EPS) from P. kribbensis PS04 had been purified and their structure was inferred to be fructosan. This study aimed to evaluate the effects of exogenous EPS treatment on plant­pathogen interactions. RESULTS: Plant defense genes such as phenylalanine ammonia-lyase, catalase, chitinase, allene oxide synthase, and PR1a proteins were significantly induced by exogenous EPS treatment. Moreover, subsequent challenge of EPSpretreated plants with the pathogens (R. solani or tobacco mosaic virus) resulted in higher expression of defenseassociated genes. Increased activities of defense-associated enzymes, total phenols, and flavonoids were also observed in EPS pretreated plants. The contents of malondialdehyde in plants, which act as indicator of lipid peroxidation, were reduced by EPS treatment. CONCLUSIONS: This study comprehensively showed that EPS produced from P. kribbensis PS04 enhances disease resistance in plants by the activation of defense-associated genes as well as through the enhancement of activities of defense-related enzymes.

Paenibacillus dendritiformis DDS2: A new member to alkaline-thermophilic protease producers

@#Aim: Most of the industrial processes require thermostable alkaline proteases. Thus, a search was initiated to isolate and characterize a bacterium which can produce thermostable alkaline protease. Methodology and results: Best higher titer thermostable alkaline protease producing wild type organism was screened from beef, dog and fish decaying soil samples. Among the 92 bacterial strains, three strains which produced alkaline proteases having activities at pH 10.5 and above 70 °C were selected. Among the three strains, the one from the dog decaying soil (Strain DDS2 ; DDS-dog decaying soil) which produced the protease showing highest activity at pH 10.5 and 73 °C and stability (half-life: 10.5 h) without additives was selected and identified. Based on the biochemical and morphological studies, strain DDS2 could be either Paenibacillus dendritiformis or P. thiaminolyticus. From 16S rDNA sequencing, the strain DDS2 was confirmed as P. dendritiformis. Conclusion, significance and impact: This is the first report published to show that P. dendritiformis is a protease producer and the organism was named as P. dendritiformis DDS2.

Isolation and Characterisation of Endophytic Strain Paenibacillus polymyxa SR19 from Urtica dioica and the Study of Their Effect against Fusarium oxysporum f. sp Tomato

In an attempt to obtain biological control agents for fusariose wilt, a total of 54 endophytic bacterial strains were isolated from the root of plants Urtica dioica and screened for in vitro antagonist activity against Fusarium oxysporum. Among the 54 bacterial isolates, 27 isolates exhibited more than 60% inhibition of mycelia growth of Fusarium oxysporum. The strain R19 which exhibited the most obvious antagonistic activity was selected for greenhouse studies. The SR19 had no pathogenicity and was identified as Paenibacillus polymyxa based on its phenotypical and biochemical properties as well as its 16S rRNA gene sequence. Growth chamber studies resulted in statistically significant increases in inoculating tomato seedling with the endophytic strain SR19 which in turn resulted in improving plant seedling stand by 32% and increasing fresh weights of root and fresh weight of aerial biomass of plants over the untreated pathogen control by 6.95 g and 7.96 g, respectively. Strain SR19 is a potential biological control agent that may contribute to the protection of tomato plants against fusariose wilt.

Production optimization, purification, expression, and characterization of a novel α-L-arabinofuranosidase from Paenibacillus polymyxa

Background: α-L-Arabinofuranosidase (EC 3.2.1.55) catalyzes the hydrolysis of terminal α-L-1,2-, -1,3-, and -1,5- arabinofuranosyl residues in arabinose-containing polymers, and hence, it plays an important role in hemicellulose degradation. Herein, the bacterium Paenibacillus polymyxa, which secretes arabinofuranosidase with high activity, was selected for enzyme production, purification, and characterization. Results: Medium components and cultural conditions were optimized by the response surface method using shake flask cultures. Arabinofuranosidase production reached 25.2 U/mL under optimized conditions, which were pH 7.5, 28°C, and a basic medium supplemented with 1.5 g/L mannitol and 3.5 g/L soymeal. Furthermore, the arabinofuranosidase secreted by P. polymyxa, named as PpAFase-1, was partially purified from the supernatant using a DEAE Sepharose Fast Flow column and a hydroxyapatite column. The approximate molecular mass of the purified PpAFase-1 was determined as 56.8 kDa by SDS-PAGE. Protein identification by mass spectrometry analysis showed that the deduced amino acid sequence had significant similarity to the glycosyl hydrolase family 51. The deduced gene of 1515 bp was cloned and expressed in Escherichia coli BL21 (DE3) cells. Purified recombinant PpAFase-1 was active toward p-nitrophenyl-α-L-arabinofuranoside (pNPAraf). The Km and kcat values toward pNPAraf were 0.81 mM and 53.2 s−1 , respectively. When wheat arabinoxylan and oat spelt xylan were used as substrates, PpAFase-1 showed poor efficiency. However, a synergistic effect was observed when PpAFase-1 was combined with xylanase from Thermomyces lanuginosus. Conclusion: A novel GH51 enzyme PpAFase-1 was cloned from the genome of P. polymyxa and expressed in E. coli. This enzyme may be suitable for hemicellulose degradation on an industrial scale.

Promotion of iron nutrition and growth on peanut by Paenibacillus illinoisensis and Bacillus sp. strains in calcareous soil

ABSTRACT This study aimed to explore the effects of two siderophore-producing bacterial strains on iron absorption and plant growth of peanut in calcareous soil. Two siderophore-producing bacterial strains, namely, YZ29 and DZ13, isolated from the rhizosphere soil of peanut, were identified as Paenibacillus illinoisensis and Bacillus sp., respectively. In potted experiments, YZ29 and DZ13 enhanced root activity, chlorophyll and active iron content in leaves, total nitrogen, phosphorus and potassium accumulation of plants and increased the quality of peanut kernels and plant biomass over control. In the field trial, the inoculated treatments performed better than the controls, and the pod yields of the three treatments inoculated with YZ29, DZ13, and YZ29 + DZ13 (1:1) increased by 37.05%, 13.80% and 13.57%, respectively, compared with the control. Based on terminal restriction fragment length polymorphism analysis, YZ29 and DZ13 improved the bacterial community richness and species diversity of soil surrounding the peanut roots. Therefore, YZ29 and DZ13 can be used as candidate bacterial strains to relieve chlorosis of peanut and promote peanut growth. The present study is the first to explore the effect of siderophores produced by P. illinoisensis on iron absorption.

Characterization and comparison of phytase production by Bacillus and Paenibacillus strains from Thai soils

Aims: The objective of this research was to isolate, screen and identify phytase-producing bacteria from soils and a potent isolate was selected for its phytase production. Methodology and results: Eight spore-forming bacteria isolated from agricultural soils in Thailand were screened for their phytase production. They were identified as Bacillus and Paenibacillus strains based on their phenotypic characteristics and 16S rRNA gene sequence analyses. The phytase production by Bacillus amyloliquefaciens CH3-1 [Group I(a)] was 20.956 ± 0.099 U/mL, while Bacillus subtilis SR9-3 [Group I(b)] produced 20.588 ± 0.099 U/mL. Five isolates in Group I(c), identified as Bacillus aryabhattai, produced phytase at levels ranging from 2.436 ± 0.116 to 20.910 ± 0.000 U/mL, while Paenibacillus cineris CM5-3 (Group II) produced 1.261 ± 0.111 U/mL. A potent strain, CH3-1, produced the highest phytase when cultivated in Phytate Specific Medium (PSM) supplemented with 1% glucose, at pH 7.0 and incubated at 45 °C. Additionally, wheat bran and sorghum seed (0.5%) substrates were used to induce phytase production by replacing Na-phytate. Conclusion, significance and impact of study: Phytase producing bacteria were isolated from soils in Thailand. Gram-positive spore forming thermotolerant Bacillus strains displayed higher phytase activity than a Paenibacillus strain. A potent strain, CH3-1, could utilize agricultural waste as a substrate, which may be useful for animal feed supplementation.

Adherence of Streptococcus pneumoniae and expression analysis of neuraminidase gene (NanA and NanB) after interaction of A549 human lung epithelial cells with pneumococcal strains of various serotypes

Aims: Streptococcus pneumoniae is the most common bacterial respiratory pathogen that can lead to invasive diseases such as pneumonia, bacteremia, and meningitis. The interaction of S. pneumoniae with host respiratory epithelial cells is crucial in the colonization of human respiratory tract and involve in the virulence. The aim of the study is to investigate the adherence of S. pneumoniae and the effect of serotypic variation on neuraminidase genes (NanA and NanB) after interaction of A549 human lung epithelial cells with S. pneumoniae serotypes. Methodology and results: Six different serotypes of S. pneumoniae were used (1, 3, 5, 19F, 23F, and 14). A549 human lung epithelial cells were inoculated with pneumococcal strains of different serotype for 3 hours. The number of adherent bacteria was determined by serial dilution followed by spread plate technique on tryptic soy agar supplemented with 5% sheep blood. Bacterial RNA was harvested from the infected A549 cells. The differential expression level of neuraminidases was observed by quantitative real-time PCR (qRT-PCR). Based on bacterial adherence assay, serotype 14 showed highest adherence, meanwhile, serotype 23F showed lowest adherence. This suggests that serotype 14 has a better affinity to adhere to A549 cells as compared to serotype 23F. Higher NanA gene expression was observed in serotype 5, 23F and 19F, while lower expression in serotype 14. In contrast, NanB gene shows low-level expression in serotype 23F and 19F, while higher expression in serotype 14. This postulates that NanA and NanB gene may have different functions in the pathogenesis of S. pneumoniae. Conclusion, significance and impact of study: Our finding on differential expression of neuraminidase gene of S. pneumoniae of various serotypes on A549 cells might give a better understanding of host pathogen interaction between bacteria serotypes and host cell.

Five Cases of the Gram Variable Bacterium Paenibacillus urinalis Isolated from Clinical Specimens and its Clinical Significance

Paenibacillus urinalis was first isolated from the urine of a woman in 2008, and was reported to be a contaminant. Here, we report 5 cases of P. urinalis isolated over 5 months at a tertiary hospital. Using an API kit, 4 cases were classified as Cellulomonas species. Owing to the low reliability of API kit results and Gram stain results indicating gram variable bacilli for few specimens, MALDI-TOF MS and 16S rRNA gene sequencing were performed for identification. The last case showed Gram variable bacilli, and therefore, based on previous experience, 16S rRNA gene base sequence analysis was carried out without an additional API kit. All isolated strains were confirmed to be P. urinalis, and were judged to be contaminants. As for Gram variable bacteria, the use of current biochemical identification systems may lead to misidentification as other bacteria, which may cause unnecessary or improper use of antibiotics. Moreover, whereas most of the Paenibacillus species are reported to be contaminants, some of them are being reported as sources of infection. Therefore, more accurate identification will be necessary in the future. Accordingly, it is expected that accurate identification of this genus will help clinical physicians make decisions regarding appropriate treatment and use of antibiotics.

Microbial-induced remediation of Zn2+ pollution based on the capture and utilization of carbon dioxide

Background: Microbial-induced remediation of Zn2+ pollution based on the capture and utilization of carbon dioxide was investigated. In this study, carbon dioxide was absorbed and transformed into carbonate ions under the enzymatic action of Paenibacillus mucilaginosus, which was being utilized to mineralize Zn2+. Results: The compositional and morphological properties of the precipitations were studied using Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The thermal properties of the precipitates were investigated by thermogravimetric-differential scanning calorimetry (TG-DSC). The FTIR results confirmed that the functional groups of the precipitates were CO3² − and OH−. The XRD and EDS patterns showed that basic zinc carbonate could be obtained successfully by Microbial-induced remediation. The SEM micrographs demonstrated that the precipitates were in the nanometer range with sizes of 100-200 nm and were sphere-like in shape. Conclusions: The TG-DSC results showed that weight loss of the precipitates occurred around 253°C. The FTIR and TG-DSC results were in accord with the XRD and EDS results and proved again that the precipitates were basic zinc carbonate. This work thus demonstrates a new method for processing Zn2+ pollution based on the utilization of carbon dioxide.

Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters

Abstract A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth.

Characterization of lignocellulolytic bacterial strains associated with decomposing wood residues in the Lagos lagoon, Nigeria

Aims: The presence of lignocelluloses, especially sawdust in the Lagos lagoon and the attendant ecological problems warranted studies on their degradation. This study aimed to isolate and identify the indigenous bacterial strains capable of utilizing lignocellulosic wastes under the prevalent tropical estuarine conditions. Methodology and results: Nine bacterial species were obtained by elective culture from decomposing wood residues in the lagoon. They were identified on the basis of morphology, biochemical characteristics and analysis of their 16S rRNA gene sequences as Streptomyces, Bacillus and Paenibacillus species. They were cultured on various ligninrelated lignocellulosic substrates over a period of 7 to 12 days. All the isolates showed moderate to very good growth on sugarcane baggase. Streptomyces albogriseolus strain AOB and Paenibacillus sp. ROB showed good growth on grass while on sawdust, only Streptomyces AOB, and Bacillus megaterium strain NOB showed good growth. High performance liquid chromatographic analysis showed that the Streptomyces species completely utilized coniferyl alcohol, B. megaterium strain NOB utilized 90-100% of all the lignin- related aromatic compounds. All the bacterial species utilized less than 40% of sinapyl alcohol, Bacillus sp. OOB and Paenibacillus sp. strain ROB failed to utilize vanillic acid. Conclusion, significance and impact of study: The isolates degraded lignocellulosic wastes and lignin-related compounds. The role of fungi in the breakdown of lignocellulose in the Lagos lagoon had been the subject of previous research considerations whilst the role of bacteria spp was unreported. Autochthonous bacterial species may equally play a role in the bio-rehabilitation of the sawdust-polluted water of the Lagos lagoon. Keywords: Lignocellulose, Streptomyces, Bacillus, Paenibacillus, pollution

Isolation and characterization of cellulose degrading ability in Paenibacillus isolates from landfill leachate

Aims: Cellulases are enzymes that convert cellulose into glucose molecules, and are produced by various microorganisms in the environment. Due to their importance to the biofuel industry, there is a need to screen for more efficient varieties of cellulases. In this study, leachate samples from a landfill site were screened for cellulolytic bacteria. Methodology and results: Leachate samples obtained from a landfill collection pond were cultured in an enriched cellulose medium. Two cellulolytic isolates, designated MAEPY1 and MAEPY2, were isolated and further characterized. Phenotypic profiles and phylogenetic analyses using sequences of 16S rRNA, gyrB and whole genome suggested that these isolates are new strains of the Paenibacillus genera. The crude enzyme extracts from both isolates have cellulose degradation activity at approximately 0.1-0.2 IU/mg under working conditions of pH 6 and 55 °C. Assays using other lignocellulosic substrates showed that the crude enzyme extracts also have high xylan degradation activity. Conclusion, significance and impact of study: Paenibacillus sp. are known to produce multiple enzymes for lignocellulolytic degradation and the present results suggest that isolates described in this study, MAEPY1 and MAEPY2, are excellent candidates deserving further study as potential producers of efficient cellulases for use in industries associated with cellulosic biomass.

A Case of Paenibacillus pasadenensis Bacteremia in a Patient with Acute Respiratory Distress Syndrome after Microsurgical Clipping

We report the first case of bacteremia by a novel Paenibacillus species, Paenibacillus pasadenensis, from a 55-year-old male patient with acute respiratory distress syndrome, following a microsurgical clipping procedure of a ruptured intracranial aneurysm. The bacterium was identified using 16S rRNA gene sequencing analysis, which was applied because current conventional methods employed in the clinical microbiology laboratory proved unsuccessful. Since this bacterium was first identified in 2006 and has never been reported elsewhere, we believe this report can provide practitioners with useful insight on the pathogenicity of this species.

Identification and characterization of the Sudanese Bacillus thuringiensis and related bacterial strains for their efficacy against Helicoverpa armigera and Tribolium castaneum.

Forty-four isolates of Bacillus thuringiensis like bacteria from various sources in different locations from Sudan were tested for their insecticidal activity. The toxicity of these isolates ranged from 6.6 to 70% to the neonates of cotton bollworm, Helicoverpa armigera at 10 ppm concentration. The most effective ones are Kb-29, St-6 and Wh-1 comparable with HD-1.  Toxicity of isolates to larvae of the red flour beetle, Tribolium castaneum ranged from 20 to 100%. Isolates St-2 and St-23 gave 100% larval mortality within 15 days of exposure and were at par with Ab-8, Ab-12, Kb-26, Kb-30, Om-4, Po-2, Po-5, Po-7, Sa-8 and Wh-5 and were also comparable with E. coli clone expressing Cry3 toxin. The most effective five isolates viz., Kb-29, St-2, St-6, St-23 and Wh-1 belonged to B. thuringiensis. The St-6 isolate, which also showed high toxicity to T. castaneum larvae, had cry1 genes along with coleopteran active cry28 genes, but not cry3 genes. Of the 25 isolates characterized with 16s DNA sequencing, seven belonged to Paenibacillus spp., one Lysinibacillus sphaericus, one Bacillus pumilus, four Bacillus spp., and rest 12 belonged to B. thuringiensis. Biochemical characterization in each species showed variation. The present study shows potential of some isolates like Kb-29, St-2, St-6, St-23 and Wh-1 as promising bioinsecticides.

Characterization of cellulase producing Bacillus and Paenibacillus strains from Thai soils.

Seventeen strains of cellulase producing bacteria were isolated from soil samples collected in Nan province, Thailand. They exhibited cellulase activity as a clear zone surrounded their colonies grown on carboxymethyl cellulose (CMC) agar medium ranged from 0.63 to 2.95 cm in diameter. Their hydrolysis capacity values were 1.65 - 7.55. The bacteria isolated were divided into 2 groups and belonged to genus Bacillus and Paenibacillus based on their phenotypic characteristics and chemotaxonomic characteristics such as meso-diaminopimelic in cell wall peptidoglycan and menaquinones of MK-7. Bacillus strains, P3-1 and P4-6 in Group I, produced maximum cellulase at 0.015 U ml-1. Their optimal pH and temperature for enzyme production and enzyme activity were 7.0 and 50 oC, respectively. The strains P3-1 and P4-6 were closely related to Bacillus velesensis LMG 22478T (100% similarity) whereas representative strain, S10-4 in Group II, was closely related to P. cellulositrophicus KCTC 13135T (98.7% similarity) based on 16S rRNA gene sequence. On the basis of their phenotypic, chemotaxonomic characteristics and phylogenetic analysis of 16S rRNA gene sequence, the strains P3-1 and P4-6 were identified as B. velesensis and the strain S10-4 was a novel species in the genus Paenibacillus.

Screening and identification of cellulase producing bacteria isolated from oil palm meal.

Oil palm (Elaeis guineensis) meal, a by-product of palm oil, is rich in fiber and contains lignocelluloses, which inhibits the absorption of the nutrients has been widely used for animal feed. The improvement of the nutrient absorption is required treating with cellulase enzyme. This study was aimed to isolate, screen and characterize the cellulase producing bacteria. Ten strains of cellulolytic bacteria were isolated from 7 oil palm meal samples collected in Phetchaburi, Prachuap Khiri Khan and Pattani provinces, Thailand. They exhibited the ability to degrade carboxymethyl-cellulose (CMC) based on the decolorization of CMC-basal agar medium using Congo red as a color indicator. They showed the cellulase hydrolysis capacity ranged from 1.56 to 4.14. All isolates were Gram positive rod-shaped bacteria and belonged to Bacillus (8 isolates), Paenibacillus (1 isolate) and Lysinibacillus (1 isolate) based on the phenotypic characteristics and 16S rRNA gene sequence analysis. Their cellulase activity ranged from 0.039±0.002 to 0.233±0.005 IU/ml when they were cultivated in broth.

Screening of endoglucanase-producing bacteria in the saline rhizosphere of Rhizophora mangle

In screening the culturable endoglucanase-producing bacteria in the rhizosphere of Rhizophora mangle, we found a prevalence of genera Bacillus and Paenibacillus. These bacteria revealed different activities in endoglucolysis and biofilm formation when exposed to specific NaCl concentrations, indicating modulated growth under natural variations in mangrove salinity.

Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil

In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant.

A Case of Paenibacillus-induced Sepsis Complicated with Pneumatocele in a Very Low Birth Weight Infant

Paenibacillus spp. are gram-positive, rod-shaped, facultative anaerobic bacteria found in nature and rarely cause diseases in humans. We report our experience with Paenibacillus-induced sepsis complicated with pneumatocele in a very low birth weight male infant with a gestational age of 29 weeks and 5 days and a birth weight of 1,380 g, who was born by cesarean section with because of preterm labor and premature rupture of membrane. On day 12 after admission, the patient presented oxygen desaturation without apnea and fever. We identified pleural effusion on chest radiography and diagnosed pneumatocele on low-dose chest computed tomography. An empirical antibiotic was administered to treat the infection. The patient's blood culture revealed gram-positive rods, and Paenibacillus spp. was identified using16s rRNA sequencing.

Production and characterization of carboxymethyl cellulase from Paenibacillus polymyxa using mango peel as substrate.

Mango peel, a solid mango processing waste, comprises 15-20% of total fruit weight. This, being a rich source of lignocelluloses, was used as substrate for carboxymethyl cellulase (CMCase) production using Paenibacillus polymyxa. Maximum CMCase production (7.814 U mg-1) was observed in a medium containing 7% mango peel (w/v) with 1.5% ammonium sulphate (w/v) at 37oC and pH 5.5. Purification to an extent of 28.24 fold was achieved by affinity column chromatography. Bands corresponding to 26.5 and 34.0 kDa molecular sizes were observed on 12% denaturing Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) while of 72 kDa on 10% non- denaturing Native-PAGE, proving its heteromeric multienzyme nature. The enzyme was stable over a range of 20-60oC and pH of 4.0-7.5. Michaelis-Menten equation constant (Km and Vmax) values of purified CMCase were 8.73 mg ml-1 and 17.805 mM ml-1 min-1, respectively.