Thermophilic Alkaline Xylanase from Newly Isolated Alkaliphilic and Thermophilic<i>Bacillus</i>sp. Strain TAR-1

Nakai, Ryuichiro; Wakabayashi, Kenji; Ishiguro, Yutaka; Aono, Rikizo; Horikoshi, Koki

doi:10.1271/bbb.58.78

Cited by 69 publications

(39 citation statements)

References 6 publications

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“…2). Optimal activity near neutral pH has been described for many bacterial xylanases [24,28]. The optimal temperature for Xyl53 activity was found to be 55°C, although the enzyme exhibited 90% of its activity in the range from 50 to 60°C (Fig.…”

Section: Puriwcation Of Xyl36 Xyl53 and Zymogram Analysismentioning

confidence: 90%

Purification and characterization of two sugarcane bagasse-absorbable thermophilic xylanases from the mesophilic Cellulomonas flavigena

Santiago-Hernández

Vega-Estrada

Montes-Horcasitas

et al. 2007

J Ind Microbiol Biotechnol

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We report the purification and characterization of two thermophilic xylanases from the mesophilic bacteria Cellulomonas flavigena grown on sugarcane bagasse (SCB) as the only carbon source. Extracellular xylanase activity produced by C. flavigena was found both free in the culture supernatant and associated with residual SCB. To identify some of the molecules responsible for the xylanase activity in the substrate-bound fraction, residual SCB was treated with 3 M guanidine hydrochloride and then with 6 M urea. Further analysis of the eluted material led to the identification of two xylanases Xyl36 (36 kDa) and Xyl53 (53 kDa). The pI for Xyl36 was 5.0, while the pI for Xyl53 was 4.5. Xyl36 had a Km value of 1.95 mg/ml, while Xyl53 had a Km value of 0.78 mg/ml. In addition to SCB, Xyl36 and Xyl53 were also able to bind to insoluble oat spelt xylan and Avicel, as shown by substrate-binding assays. Xyl36 and Xyl53 showed optimal activity at pH 6.5, and at optimal temperature 65 and 55 degrees C, respectively. Xyl36 and Xyl53 retained 24 and 35%, respectively, of their original activity after 8 h of incubation at their optimal temperature. As far as we know, this is the first study on the thermostability properties of purified xylanases from microorganisms belonging to the genus Cellulomonas.

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Section: Puriwcation Of Xyl36 Xyl53 and Zymogram Analysismentioning

confidence: 90%

Purification and characterization of two sugarcane bagasse-absorbable thermophilic xylanases from the mesophilic Cellulomonas flavigena

Santiago-Hernández

Vega-Estrada

Montes-Horcasitas

et al. 2007

J Ind Microbiol Biotechnol

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“…As given in Fig. 2 the enzyme showed optimum activity at 50 "C. Reports on the alkaline xylanases that showed higher temperature optima were scarce (NAKAMURA et al 1994).…”

Section: Effect Of Temperature On Activity and Stability Of Xylanasementioning

confidence: 97%

“…Endoxylanases [ 1,4-P-D-xylan xylanohydrolases, (E.C.3.2.1.8)] hydrolyse xylan to xylooligosaccharides and xyloses while P-xylosidases [ 1,4-/3-D-xylan xylohydrolases (E.C.3.2.1.37)l catalyse the release of xylosyl residues by terminal attack of xylooligosaccharides (DAHLBERG et al 1993, SCHOFIELD et al 1993, NAKAMURA et al 1994. The most important application of xylanases is the pretreatment of pulps prior to bleaching in pulp and paper industry.…”

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confidence: 99%

Isolation and screening for alkaline thermostable xylanases

Subramaniyan

Prema

Ramakrishna

1997

J. Basic Microbiol.

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Screening of xylanase producing microorganisms from soil samples and hemicellulosic materials were carried out in three stages using wheat bran extract agar medium, xylan agar medium and liquid medium with xylan. One of the isolates, identified as BaciZlus SSP-34 produced 100 times more xylanase activity than the other isolates. Maximum enzyme production by this isolate was observed after 102 hours of growth. The optimum pH for the crude xylanase preparation range from 6-8 and optimum temperature was at 50 "C. The enzyme was stable up to 20 minutes at 50 "C and at pH range 4.5-9.0 for more than 2 hours.Xylans are the second most abundant carbohydrates next to cellulose having a linear backbone of /3-1,4 linked xyloses and xylanases found commonly in microorganisms catalyse the hydrolysis of xylans. Endoxylanases [ 1,4-P-D-xylan xylanohydrolases, (E.C.3.2.1.8)] hydrolyse xylan to xylooligosaccharides and xyloses while P-xylosidases [ 1,4-/3-D-xylan xylohydrolases (E.C.3.2.1.37)l catalyse the release of xylosyl residues by terminal attack of xylooligosaccharides (DAHLBERG et al. 1993, SCHOFIELD et al. 1993, NAKAMURA et al. 1994. The most important application of xylanases is the pretreatment of pulps prior to bleaching in pulp and paper industry. The hydrolysis of xylan facilitates the release of lignin from paper pulp and hence reduces the usage of chlorine as the bleaching agent (SHOHAM et al. 1992). Thermostable xylanases with alkaline pH range will be more efficient in kraft pulping process due to the higher temperatures and pH ranges necessary for wood chip cooking (SHOHAM et al. 1992, VIIKARI et al. 1994. Only certain bacterial and a few actinomycete xylanases have been reported earlier with pH optima in the neutral or alkaline ranges (NAKAMURA et al. 1993). In search of novel thermostable alkaline xylanases suitable for industrial applications, studies have been initiated on isolating microorganisms that produce xylanases. The following paper reports the isolation and screening of potent xylanase producing microorganisms and primary characterisation of the xylanases from the selected isolate. Materials and methodsIsolation and screening: Forest soil samples from Kallar and Ponmudi, Kerala (India) and various hemicellulose containing substrates (wheat bran, bagasse and rice straw) exposed to atmosphere were suspended in sterile distilled water. Suspensions after serial dilution were spread on wheat bran extract agar plates containing (gfl): wheat bran, 50.0; peptone, 5.0; NaCI, 5.0; yeast extract, 3.0 and agar, 20.0. In secondary screening, these cultures were spread on xylan agar plates using 0.5% xylan (Oat spelts xylan from SIGMA Chemicals Co.) instead of wheat bran. After six days of incubation, colonies that showed areas of clear zones with a radius of a minimum of 1 cm were selected for further screening in liquid medium where oat spelts xylan was the sole carbon source. Xylan liquid culture medium (NAKAMURA et al. 1993) contained (gfl): xylan, 5.0; peptone, 5.0; yeast extract, 5.0; K2HP04, 1.0 and M...

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“…Kondisi pH optimum dibutuhkan oleh enzim untuk mengaktifkan seluruh enzim yang mengikat substrat dan mengubahnya menjadi produk. Hasil ini hampir sama dengan hasil penelitian Nakamura et al (1994), pada xilanase dari isolat Bacillus sp. TAR-I alkalofilik dan termofilik yang memperlihatkan aktivitas xilanase pada kisaran pH 5-9,5 dan optimum pada pH 9.…”

Section: Karakterisasi Xilanase Penentuan Ph Dan Suhu Optimumunclassified

Isolasi Identifikasi Bakteri Penghasil Xilanase serta Karakterisasi Enzimnya

Richana¹,

Irawadi²,

Nur³

et al. 2016

J. AgroBiogen

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Isolation and Identification of Xylanase Producing Bacteria and Characterization of Its Enzyme Properties. Nur Richana, Tun T. Irawadi, Anwar Nur, and Khaswar Syamsu. Xylanase is an extracellular enzyme produced by microorganisms. This enzyme is able to hydrolise xylane (hemicellulose) to produce xylooligosaccharide and xylose. Thermoalkaliphilic xylanase is an agent that can be used as a substitute in the pulp whitening process instead of chlorine. A study was done to isolate, identificate of bacteria and characterize xylanase. The isolation of xylanase producing bacteria has been done from soil and waste of starch industry. Colonies which produced clearing zone were presumed as xylanolytic bacteria and chosen for further screening. Identification of potential isolate in xylanase production was done using 16S ribosomal RNA sequencing. Isolate Bacillus pumilus RXA-III5 originated from lime or alkaline soil was more potential isolate in xylanase production than other 24 isolates. Precipitation of xylanase, that was done using ammonium sulphate followed by dialyzes produced xylanase of a higher specific activity (267.1 U.mg ). Xylanase was done at purification produced three fractions of xylanase. Xylanase characteristics consist of pH and temperature (9 and 50 o C), K m and V maks value 6 mg.ml -1 and 0.2 mol.minute -1 , respectively. The Fe 2+ was the strongest activetor and Mg 2+ was the strongest inhibitor activity. This enzyme was detected as a cellulose-free xylanase. Xylanase is a prospective agent for bio-bleaching of paper.Key words: Isolation, identification, bacteria, xylanase. PENDAHULUANXilanase merupakan enzim ekstraseluler yang dapat menghidrolisis xilan (hemiselulosa) menjadi xilo-oligosakarida dan xilosa. Xilanase dapat dihasilkan oleh mikroba melalui proses fermentasi. Aplikasi xilanase untuk industri di antaranya untuk industri pangan, pakan, dan pemutih bubur kertas/pulp. Penggantian penggunaan klor dengan enzim xilanase untuk pemutihan pulp telah memberikan peluang untuk aplikasi bioteknologi dan sekarang telah digunakan pada beberapa pabrik kertas (Bourbonnais et al. 1997, Beg et al. 2000. Hak Cipta © 2008, BB-BiogenXilanase dapat dihasilkan oleh mikroba melalui proses fermentasi, yang biasanya dihasilkan oleh bakteri atau khamir. Untuk pembuatan kertas diperlukan xilanase yang bersifat termostabil dan tahan pada pH alkali. Xilanase komersial untuk proses pemutihan pulp telah mulai dipasarkan. Namun demikian semua enzim komersial ini masih belum memenuhi kriteria ideal yang dibutuhkan untuk aktivitas enzimatik yang diperlukan, yaitu aktivitas optimum pada pH 10 dan suhu lebih dari 90 o C (Kulkarni dan Rao 1996). Oleh karena itu, masih diperlukan upaya untuk mencari galur mikroba unggul yang tahan pada pH dan suhu tinggi (alkalofilik termofilik), atau setidaknya tahan pada pH tinggi.Upaya pemanfaatan enzim dari mikroba potensial hasil isolasi diperlukan pengetahuan mengenai karakteristik dari enzim yang dihasilkan. Pencirian suatu enzim di laboratorium, dilakukan dengan menetapkan kin...

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Thermophilic Alkaline Xylanase from Newly Isolated Alkaliphilic and ThermophilicBacillussp. Strain TAR-1

Cited by 69 publications

References 6 publications

Purification and characterization of two sugarcane bagasse-absorbable thermophilic xylanases from the mesophilic Cellulomonas flavigena

Purification and characterization of two sugarcane bagasse-absorbable thermophilic xylanases from the mesophilic Cellulomonas flavigena

Isolation and screening for alkaline thermostable xylanases

Isolasi Identifikasi Bakteri Penghasil Xilanase serta Karakterisasi Enzimnya

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