Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge / Modulando a produção de xilanase por Bacillus pumilus BS131 por meio da otimização usando lodo de fibra residual

Abstract In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.

Resumo Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.

Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge / Modulando a produção de xilanase por Bacillus pumilus BS131 por meio da otimização usando lodo de fibra residual

In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.

Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.

Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge

Abstract In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.

Resumo Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.

Selection of microorganisms and preparation of inoculants with potential applications in the remediation of soil contaminated with hexazinone / Seleção de microrganismos e preparo de inoculantes com potencial de aplicação na remediação de solo contaminado com hexazinona

ABSTRACT: The excessive use of agrochemicals negatively impacts the environment, making the development of sustainable technologies for the reduction of contaminants in soil necessary. Hexazinone is the herbicide most used for sugarcane crops and persists in the environment. Moreover, its main route of degradation in the soil is through microorganisms. Therefore, six microorganisms were selected that presented growth in the presence of the herbicide; SCR1 - Microbacterium arborescens; SCR2 - Bacillus pumilus; SCM3 - Stenotrophomonas maltophilia; SCM4 - Bacillus cereus; SCM5A - M. arborescens; and SCM5B - B. safensis. A test was performed to evaluate the ability of each lineage in phosphate solubilization. For the Ca3(PO4)2 solubilization test, the strains that showed the best results were B. pumilus and S. maltophilia. Subsequently, the inoculants were prepared and the concentrations after plating were 2.71 × 109 CFU mL-1 for B. pumilus, 1.02 × 109 CFU mL-1 for S. maltophilia, and 1.14 × 1010 CFU mL-1 for a combination of the two strains. These were satisfactory values for use as inoculants.

RESUMO: O uso de agroquímicos resulta em impactos ambientais e torna-se necessário o emprego de tecnologias sustentáveis para diminuição de contaminantes no solo. O hexazinona é o herbicida mais utilizado para a cultura da cana-de-açúcar, e apresenta persistência no ambiente. A principal via de degradação no solo é por meio de microrganismos. Com isso, selecionou-se seis microrganismos que apresentaram crescimento na presença do herbicida: SCR1 - Microbacterium arborescens; SCR2 - Bacillus pumilus; SCM3 - Stenotrophomonas maltophilia; SCM4 - Bacillus cereus; SCM5A - M. arborescens; SCM5B - Bacillus safensis. Foi realizado um teste para avaliar a habilidade de cada linhagem na solubilização de fosfatos, e no caso da solubilização de Ca3(PO4)2, as linhagens que apresentaram melhores resultados foram B. pumilus e S. maltophilia. Posteriormente, os inoculantes foram preparados e a concentração após plaqueamento de 2,71x109 UFC mL-1 para B. pumilus, 1,02x109 UFC mL-1 para S. maltophilia e consórcio com as duas linhagens 1,14x1010 UFC mL-1 apresentaram valores satisfatórios para utilização como inoculantes.

Use of bacterial endospore with longer shelf-life in improved retting of jute

Aim: The aim of the present study was to evaluate the retting of jute (Corchorus olitorius L. and C. capsularis L.) using the endospores of microbial consortium of three strains of Bacillus pumilus with extended shelf-life. Methodology: Endospore and vegetative cells of Bacillus pumilus were tested for viability by introducing them into different temperature, pH, UV radiation and antibiotics. Laboratory, as well as field-trials of jute retting was performed with 6 and 18-months-old endospores and vegetative cells of Bacillus pumilus with estimation of enzymatic activities for comparison of their retting efficiency. Results: Endospores of Bacillus pumilus recorded very high colony forming unit (109 to 108ml-1) compared to their vegetative cells (106 to 104ml-1) after 6 to 18 months of their preservation. Endospores also showed higher resistance to temperature, pH, UV irradiation and antibiotic than their vegetative forms. High colony forming unit and higher release of pectinolytic and xylanolytic enzymes during retting of jute by endospores resulted in complete of jute retting in 10 days with good quality jute fibre compared to talc based formulation. Interpretation: It can be concluded from the study that endospores remained highly efficient in rejuvenating higher CFU and quantitatively larger pool of enzymes to accelerate retting of jute after prolonged preservation. Therefore, the endospores of Bacillus pumilus can be used cost effectively in place of their talc based formulation for higher shelf life of the product, faster retting and better fibre quality of jute.

Biodegradación de xantano por Bacillus pumilus / Biodegradation of xanthan gum by Bacillus pumilus

La goma xantano es utilizada en la industria hidrocarburífera para la extracción terciaria del petróleo. En su utilización en la cuenca del Golfo San Jorge, se observaron inconvenientes con la perdida de estructura del gel que se forma. Se buscó el agente causal de la perdida de estructura del gel. Se aisló e identifico a Bacillus pumilus como la bacteria capaz de crecer utilizando la goma xantano como única fuente de carbono y energía, modificando la viscosidad del gel y así cambiando sus propiedades

Xanthan gum is used in the oil industry for the tertiary extraction of petroleum. In its use in the basin of the Gulf of San Jorge, there were problems with the loss of gum structure. Strain identified by Bacillus pumilus was isolated from oil water industry and was grown in the medium with xanthan as carbon source. This change modifies the viscosity of the gum and thus changing its properties

Degradation of monoaromatics by Bacillus pumilus MVSV3

ABSTRACT Monoaromatics, such as benzene, toluene, ethylbenzene and xylene (BTEX), are simple aromatic compound that are highly toxic due to their high solubility nature. Many chemical and physical methods for their degradation and breakup into nontoxic products are available, but still use of microorganism is preferred over these processes. In this present study Bacillus pumilus MVSV3 (Accession number JN089707), a less explored bacteria in the field of BTEX degradation, isolated from petroleum contaminated soil is utilized for BTEX degradation. At optimized conditions the isolate degraded 150 mg/L of BTEX completely within 48 h. GC-MS analysis revealed that the microorganism produces catechol and muconic acid during degradation indicating an ortho pathway of degradation. Enzyme assays were carried out to identify and characterize catechol 1, 2- dioxygenase (C12D). The optimal temperature and pH for the enzyme activity was identified as 35 (C and 7.5, respectively. SDS-PAGE revealed the molecular weight of the enzyme to be approximately 35,000 Da. Zymography analysis indicated the presence of three isoforms of the enzyme. Hence Bacillus pumilus MVSV3 and the isolated C12D, proved to be efficient in degrading the toxic aromatic compounds.

Screening and Production of Biodegradable Polyester Poly(3-hydroxybutyrate) by Bacteria Endophytic to Brassica nigra L.

Aims: Poly(3-hydroxybutyrate) [P(3HB)], the microbially produced biodegradable thermoplastics has find wide range of applications in recent years. Development of low cost production strategies utilizing novel organisms is a crucial challenge. Present study is aimed to isolate and screen bacterial endophytes of Brassica nigra L. for the production of P(3HB). Place and Duration of Study: The experiments were performed in the Microbiology Laboratory, Department of Botany, University of Calcutta, Kolkata during 2012-2014. Methodology: Culturable bacterial endophytes were isolated from surface sterilized healthy tissues of B. nigra L. and screened for P(3HB) production in mineral salts medium. The chloroform extracted dried polymer was treated with H2SO4 and quantified spectrophotometrically at 235 nm. Results: About 78% of the bacterial endophytes recovered from surface sterilized B. nigra L. tissues showed different degrees of P(3HB) accumulation. Isolates (9) showing P(3HB) accumulation exceeding 10% of the cell dry weight (CDW) were characterized and tentatively identified as members of Bacillus, Pseudomonas, Xanthomonas, Alcaligenes and Acetobacter. The most potent isolate, BNL 06 identified as Bacillus pumilus BNL 06 (GenBank Accession No. KP202723), accumulated P(3HB) accounting 18% of CDW with an yield of 0.55 g/l. Finally the nature of the polymer was further confirmed by FTIR analysis. Conclusion: Exploration of the endophytic bacterial diversity of B. nigra L. have clearly revealed the potential of Bacillus pumilus BNL 06 for P(3HB) production as an alternative source of thermoplastics.

Response Surface Methodology Based Optimization of a New Isolate Bacillus pumilus ZR LS S2 with Fibrinolytic Activity.

Aims: The work aims to isolate potential fibrinolytic enzyme producing isolates from various samples with the aim of developing a suitable optimization strategy using response surface methodology. Study Design: Plackett- Burman and Face centered central composite design was used. Place and Duration of Study: Department of Life Sciences, University of Calicut. Methodology: Various samples were screened using fibrin agar plates for the isolation of fibrinolytic enzyme producing bacteria and was identified by 16 s rRNA sequencing. Further the physical parameters and media components were optimized using Plackett- Burman design and face centered central composite design. Results: A novel isolate with fibrinolytic potential was isolated and was identified as Bacillus pumilus ZR LS S2. A novel media was formulated using this data, for the production of fibrinolytic enzyme, which contains peptone, casein, MgSO4 and NaCl. The isolate produced 6738.384 U/ml of enzyme in optimized conditions. Conclusion: Initial studies were performed for the production of the fibrinolytic enzyme by the novel isolate, further studies are required to effectively validate the potential of the fibrinolytic enzyme and to develop effective application for the same.

Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2

Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The lip gene consisted of 650 bp. The experimental molecular weight of SG2 lipase was nearly double that of its theoretical molecular weight, thus suggesting the existence of the functional lipase as a covalent dimer. The proteolytic cleavage sites of the lipase would have been made inaccessible by dimerisation, thus rendering the lipase resistant to protease.

Properties of a biosurfactant produced by Bacillus pumilus using vinasse and waste frying oil as alternative carbon sources

Biosurfactants are chemical molecules produced by the microorganisms with potential for application in various industrial and environmental sectors. The production parameters and the physicochemical properties of a biosurfactant synthesized by Bacillus pumilus using different concentrations of vinasse and waste frying oil as alternative carbon sources were analyzed. The microorganism was able to grow and produce a biosurfactant using both the residues. The surface tension was reduced up to 45 mN/m and the maximum production of crude biosurfactant was 27.7 and 5.7 g/l for vinasse and waste frying oil, respectively, in concentration of 5%. The critical micelle concentration (CMC) results of 1.5 and 0.2 g/l showed the efficiency of the biosurfactant produced on both the substrates. The results showed that the alternative substrates could be used for the production of an efficient biosurfactant by B. pumilus. These properties have potential for industrial and environmental applications.

Studies on degradation efficiency of polycaprolactone by a naturally-occurring bacterium.

We report a strain of Bacillus, isolated from the rhizosphere of the mangrove Sesuvium portulacastrum, that degrades polycaprolactone (PCL) on timescales that are a factor of three shorter than hitherto reported, with complete degradation in only 20 days. The bacterium has been identified as Bacillus pumilus by means of 16S rRNA gene sequencing and FAME analysis; it secretes proteases and lipases and its ‘de-polymerase’ activity is evident by the zone of clearing in emulsified PCL. It is an aerobic chemoheterotroph capable of utilizing a variety of carbohydrates. Although not a true psychrophile, is a mesophile, growing optimally over a temperature range 30-45° C and pH range 5-12.5. It is a halophile tolerating NaCl concentrations up to 10% w/v, and is unique in degrading and utilizing PCL and its monomer, e-caprolactone (CL), as a sole carbon source. Degradation of PCL was monitored using Fourier Transform Infrared (FTIR) spectroscopy, light microscopy and scanning electron microscopy (SEM). This degradation was found to be enhanced by salts (NaCl, KCl, MgSO4, Na2HPO4) and at medium pH values in excess of 7. Under the same growth conditions, another standard Bacillus pumilus strain showed somewhat reduced PCL-degradation.

Microbiological Control of Bacterial Soft Rot Caused by Bacillus pumilus Od23 on Potato.

Potato soft rot, caused by Bacillus pumilus Od23, greatly affect potato tuber quality in storage and cultivars of Mali and indicated that it can affect all potato cultivars. Bacteria isolated from soil rhizosphere samples of healthy Malian indigenous trees were screened for their antagonistic effect against this pathogen. Three actinomycetes isolates (RoN, G1P, and N1F) were the most effective microbioagents in suppressing the growth of the pathogen. The biological control essay showed the possibility of controlling potato soft rot by these three actinomycetes isolates under conservation conditions. These treatments significantly decreased soft rot compared with the untreated potato tuber slices. The microbiological control results of this study suggest that the actinomycetes isolates RoN, G1P and and J1N are effective microbioagents in controlling soft rot of potato and could be considered as promising alternative to chemical products.

Purification and partial characterization of manganese peroxidase from Bacillus pumilus AND Paenibacillus sp

The production of manganese peroxidase (MnP) from Bacillus pumilus and Paenibacillus sp. was studied under absence and presence of the inducers indulin AT, guayacol, veratryl alcohol, lignosulfonic acid and lignosulfonic acid desulfonated. Indulin AT increased the activity of B. pumilus MnP up to 31.66 U/L after 8 h, but no improve was observed for Paenibacillus sp., which reached maximum activity (12.22 U/L) after 20 h. Both MnPs produced by these microorganisms were purified in phenyl sepharose resin and the proteins from crude extracts were eluted in two fractions. However, only the first fraction of each extract exhibited MnP activities. Tests in different pH and temperature values, from pH 5.0 to pH 10.0 and 30 ºC to 60 ºC, respectively, were carried out with the purified MnP. The maximum activity reached for B. pumilus and Paenibacillus sp. MnPs were 4.3 U/L at pH 8.0 and 25 ºC and 11.74 U/L at pH 9.0 and 35 ºC, respectively. The molar masses determined by SDS-PAGE gel eletrophoresis were 25 kDa and 40 kDa, respectively, for the purified enzyme from B. pumilus and Paenibacillus sp.

Use of Bacillus pumilus CBMAI 0008 and Paenibacillus sp. CBMAI 868 for colour removal from paper mill effluent / Emprego de Bacillus pumilus CBMAI 0008 e Paenibacillus sp. CBMAI 868 para remoção da cor do efluente da indústria papeleira

Bacillus pumilus and Paenibacillus sp. were applied on the paper mill effluent to investigate the colour remotion. Inocula were individually applied in effluent at pH 7.0, 9.0 and 11.0. The real colour and COD remotion after 48h at pH 9.0 were, respectively, 41.87% and 22.08% for B. pumilus treatment and 42.30% and 22.89% for Paenibacillus sp. Gel permeation chromatography was used to verify the molar masses of compounds in the non-treated and treated effluent, showing a decrease in the compounds responsible for the paper mill effluent colour.

Bacillus pumilus e Paenibacillus sp. foram aplicados separadamente no efluente da indústria papeleira a pH 7,0, 9,0 e 11,0, para verificação da remoção da cor e da DQO. As remoções da cor real e DQO após 48h a pH 9,0 foram, respectivamente, de 41,87% e 22,08% após o tratamento com B. pumilus e 42,30% e 22,89% após tratamento com Paenibacillus sp. As massas molares dos compostos presentes no efluente não tratado e tratado foram determinadas por cromatografia de permeação em gel. O emprego dos microrganismos reduziu os compostos responsáveis pela cor do efluente da indústria papeleira.

Biochemical and molecular characterization of Bacillus pumilus isolated from coastal environment in Cochin, India / Caracterização bioquímica e molecular de Bacillus pumilus isolado do ambiente costeiro de Cochin, Índia

Bacillus species constitute a diverse group of bacteria widely distributed in soil and the aquatic environment. In this study, Bacillus strains isolated from the coastal environment of Cochin, India were identified by detailed conventional biochemical methods, fatty acid methyl ester (FAME) analysis and partial 16S rDNA sequencing. Analysis of the data revealed that Bacillus pumilus was the most predominant species in the region under study followed by B. cereus and B. sphaericus. The B. pumilus isolates were further characterized by arbitrarily primed PCR (AP-PCR), antibiotic sensitivity profiling and PCR screening for known toxin genes associated with Bacillus spp. All B. pumilus isolates were biochemically identical, exhibited high protease and lipase activity and uniformly sensitive to antibiotics tested in this study. One strain of B. pumilus harboured cereulide synthetase gene cesB of B. cereus which was indistinguishable from rest of the isolates biochemically and by AP-PCR. This study reports, for the first time, the presence of the emetic toxin gene cesB in B. pumilus.

As espécies de Bacillus constituem um grupo diversificado de bactérias amplamente distribuídas no solo e no ambiente aquático. Neste estudo, cepas de Bacillus isoladas do ambiente costeiro de Cochin, Índia, foram identificadas através de métodos bioquímicos convencionais, análise de ésteres metílicos de ácidos graxos (FAME) e sequenciamento de 16S rDNA. A análise dos dados revelou que Bacillus pumilus foi a espécie predominante na região estudada, seguido de B. cereus e B. sphaericus. Os isolados de B. pumilus foram caracterizados através da reação em cadeia da polimerase com primers arbitrários (AP-PCR), perfil de sensibilidade a antibióticos e triagem por PCR de genes de toxinas associadas com Bacillus spp. Todos os isolados de B. pumilus foram bioquimicamente idênticos, apresentaram elevada atividade de protease e lipase e foram uniformemente sensíveis aos antibióticos estudados. Um dos isolados de B. pumilus apresentou o gene cesB de B. cereus, que não foinão distinguível dos demais isolados por testes bioquímicos nem por AP-PCR. Este é o primeiro relato da presença do gene cesB da toxina eméticaem B. pumilus.

BpuAmI: a novel SacI neoschizomer from Bacillus pumilus discovered in an isolate from Amazon Basin, recognizing 5'-GAG FONT FACE=Symbol>¯ /FONT>CTC-3'

A strain of Bacillus pumilus was isolated and identified from water samples collected from a small affluent of the Amazon River. Type II restriction endonuclease activity was detected in these bacteria. The enzyme was purified and the molecular weight of the native protein estimated by gel filtration and SDS-PAGE. The optimum pH, temperature and salt requirements were determined. Quality control assays showed the complete absence of "nonspecific nucleases." Restriction cleavage analysis and DNA sequencing of restriction fragments allowed the unequivocal demonstration of 5´GAG FONT FACE=Symbol>¯ /FONT>CTC3´ as the recognition sequence. This enzyme was named BpuAmI and is apparently a neoschizomer of the prototype restriction endonuclease SacI. This is the first report of an isoschizomer and/or neoschizomer of the prototype SacI identified in the genus Bacillus.

Uma linhagem de Bacillus pumilus foi isolada e identificada de amostras de águas coletadas em um pequeno Igarapé do Rio Amazonas. Foi detectada atividade de restrição do tipo II nesta bactéria. A enzima foi purificada e o peso molecular da proteína nativa foi estimado por gel filtração e por eletroforese em gel de poliacrilamida. Foram determinados, o pH e temperatura ótimos e as necessidades de sais. Os ensaios do controle de qualidade mostraram uma ausência completa de "nucleases não específicas". As analises das clivagens e o seqüenciamento do DNA dos fragmentos de restrição permitiram uma demonstração inequívoca de que 5´GAG FONT FACE=Symbol>¯ /FONT>CTC 3´ é a seqüência de reconhecimento da enzima. Esta enzima foi denominada de BpuAmI e aparentemente é um neoesquisômero da enzima protótipo SacI. Este é o primeiro relato de um isoesquisômero e/ou neoesquisômero da enzima protótipo SacI identificada no gênero Bacillus.

The Effect of Dissolved Oxygen Tension on the Fermentation of D-ribose / 微生物学通报

After comparing the varieties of parameters in batch culture under different dissolved oxygen tension by Bacillus pumilus, we found that the demand of cell on oxygen in different phase isn' t consistent. Based on the analys is of the metabolic pathway of oxygen and the mechanism of the effect of dissolved oxygen tension on fermentation of D-ribose, a two-stage oxygen-supply control mode was proposed and experimentally proved to be available. High ribose yield and high glucose consumption rate were achieved during whole process for 44h, and the ultimate ribose concentration and cell concentration were 5.0% and 18.8% higher than former respectively.