Characterization of the recombinant xylanases in Escherichia coli from an alkaliphilic thermophilic Bacillus sp. NCIM 59

Kulkarni, Neelima; Chauthaiwale, Jyoti; Rao, Mala

doi:10.1016/0141-0229(94)00144-8

Cited by 14 publications

(9 citation statements)

References 20 publications

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“…mempunyai kisaran suhu optimum antara 50-70 o C pada pH 6-10. Kulkarni et al (1995) Yang et al (1995), aktivitas xilanase dari Bacillus sp. VI-4 mencapai optimum pada pH 9 dan suhu 55 o C. Pada pH yang sama dan suhu 60 o C, aktivitas enzim berkurang 50-20%.…”

Section: Karakterisasi Xilanase Penentuan Ph Dan Suhu Optimumunclassified

“…Hasil penelitian ini hampir sama dengan penelitian Kulkarni et al (1995) , sehingga nilai K m adalah 6 mg/ml. Berdasarkan hasil penelitian, diketahui xilanase dari B. pumilus RXA-III5 memiliki nilai V maks sebesar 0,2 mol/menit (menggunakan standar xilosa).…”

Section: Stabilitas Xilanaseunclassified

“…Hasil K m dalam penelitian ini tidak berbeda dengan hasil penelitian Kulkarni et al (1995) yang menunjukkan bahwa xilanase dari Bacillus sp. NCIM-59 menghasilkan K m 5,8-8,5 mg/ml.…”

Section: Stabilitas Xilanaseunclassified

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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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Section: Karakterisasi Xilanase Penentuan Ph Dan Suhu Optimumunclassified

Section: Stabilitas Xilanaseunclassified

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Isolasi Identifikasi Bakteri Penghasil Xilanase serta Karakterisasi Enzimnya

Richana¹,

Irawadi²,

Nur³

et al. 2016

J. AgroBiogen

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“…Ðåêîìá³íàíòí³ êñèëàíàçè, åêñïðåñîâàí³ â Escherichia coli [39] ç àëêàë³ф³ëüíîгî, òåðìîф³ëüíîгî øòàìó Bacillus sp. [35] ïðîÿâëÿëè ìåíøó ñòàá³ëüí³ñòü ïðè âèñîê³é òåìïåðàòóð³ ³ ìåíøó çäàòí³ñòü äî çâ'ÿçóâàííÿ ç êñèëàíîì â ïîð³âíÿíí³ ç êñèëàíàçàìè âèõ³äíîгî ÊCиËÀНÀÇи ÌIÊÐÎÎÐÃÀНIÇÌIÂ øòàìó, äëÿ ÿêèõ âëàñòèâî â³äñóòí³ñòü гë³êîçèëþâàííÿ. Â ñêëàä³ êñèëàíàç Talaromyces byssochlamydoides YH 50 ïðèñóòí³ çàëèøêè ìàíîçè, гëþêîçè ³ фóêîçè [62].…”

Section: продукція ксиланазиunclassified

Xylanases of Microorganisms

Varbanets¹,

Borzova²

2007

Microbiology&Biotechnology

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ÊCèËàÍàçè ÌiÊÐÎÎÐÃàÍiçÌiâÍàÿâí³ñòü âåëèêèõ ðåñóðñ³â öåëþëîçè ³ гåì³öåëþëîçè â ðîñëèíí³é á³îìàñ³ ðîáèòü âñå á³ëüø àêòóàëüíîþ ïðîáëåìó çàëó÷åííÿ ðîñëèííèõ â³äõîä³â ñ³ëüñüêîгî гîñïîäàðñòâà ³ ïðîìèñëîâîñò³ â ïðîöåñè ì³êðîáíî¿ êîíâåðñ³¿ äëÿ îòðèìàííÿ õàð÷îâîгî ³ êîðìîâîгî á³ëêà ³ ф³ç³îëîг³÷íî àêòèâíèõ ðå÷îâèí. Â³ä 20 äî 40 % â³ä ñóõî¿ âàгè ñ³ëüñüêîгîñïîäàðñüêèõ â³äõîä³â ñêëàäàº êñèëàí, ÿêèé ðàçîì ç гåì³öåëþëîçîþ óòâîðþº äðóгèé íàéá³ëüø øèðîêî ðîçïîâñþäжåíèé â á³îñфåð³ ïîë³ñàõàðèä, çäàòíèé äî â³äíîâëåííÿ. Áóëî îö³íåíî, ùî 500 ìëí òîí òàêèõ ìàòåð³àë³â ìîжóòü áóòè ùîð³÷íî äîñòóïí³ ò³ëüêè â³ä çàëèøê³â гîëîâíèõ çåðíîâèõ êóëüòóð. Òîìó ðîçâèòîê ìàëîêîøòîâíèõ òåõíîëîг³é, ÿê³ îñíîâàí³ íà гåì³öåëþëîç³, âèêëèêàº ³íòåðåñ ó äîñë³äíèê³â. йìîâ³ðíî, â íåäàëåêîìó ìàéáóòíüîìó, êñèëàí â êîìá³íàö³¿ ç öåëþëîçîþ çàáåçïå÷èòü á³ëüø³ñòü ñâ³òîâèõ ïîòðåá â ñèðîâèí³. Ìîжëèâî ïåðåäáà÷èòè, ùî â íàñòóïí³ 50 ðîê³â âóг³ëëÿ òà íåî÷èùåíà íàфòà áóäóòü çàì³íåí³ á³îìàñîþ. Â áàгàòüîõ òåõíîëîг³÷íèõ ïðîöåñàõ, ïåðø çà âñå ïðè âèðîáíèöòâ³ ïàïåðó òà öåëþëîçè, êñèëàí º íåáàжàíèì äîì³øêîì, ³ ïîâñòàº ïðîáëåìà éîгî äåгðàäàö³¿. Нåîáõ³äíî г³äðîë³çóâàòè êñèëàí òàêîж â äåÿêèõ ïðîöåñàõ õàð÷îâî¿ ³ êîðìîâî¿ ïðîìèñëîâîñò³, ïðè êîìïëåêñí³é ïåðåðîáö³ ðîñëèííèõ â³äõîä³â. Ã³äðîë³ç êñèëàíó ìîжå áóòè çä³éñíåíèé çà äîïîìîгîþ õ³ì³÷íèõ ðåàгåíò³â àáî фåðìåíòàòèâíèì øëÿõîì. Ôåðìåíòàòèâíèé г³äðîë³ç º ïåðåâàжëèâèì, îñê³ëüêè ìåíø âïëèâàº íà ñòðóêòóðó ³íøèõ âîëîêîí ³ óòâîðþºòüñÿ ìåíøå õ³ì³÷íèõ â³äõîä³â. Â õàð÷îâ³é àáî êîðìîâ³é ïðîìèñëîâîñòÿõ õ³ì³÷íèé г³äðîë³ç âçàгàë³ íåïðèéíÿòíèé. 9 Ìікробіоëоãія і біотеõноëоãія ¹ 3/2008ÊCиËÀНÀÇи ÌIÊÐÎÎÐÃÀНIÇÌIÂ Äëÿ ì³êðîáíîгî г³äðîë³çó êñèëàíó íåîáõ³äíà ä³ÿ òàêèõ фåðìåíò³â, ÿê β 1,4 êñèëàíàçè ³ β êñèëîçèäàçè. еíäî 1,4 β d êñèëàíàçà àáî 1,4 β d êñèëàí êñèëàíîг³äðîëàçà (ÊÔ 3.2.1.8) -фåðìåíò, ÿêèé çä³éñíþº íåóïîðÿäêîâàíèé г³äðîë³ç âíóòð³øíüîìîëåêóëÿðíèõ êñèëîçèäíèõ çâ'ÿçê³â êñèëàí³â àáî êñèëîîë³гîñàõàðèä³â. Âèñîêîìîëåêóëÿðí³ êñèëàíè ìàþòü ïåðåâàгó äëÿ ä³¿ фåðìåíòó â ïîð³âíÿíí³ ç îë³гîìåðíèìè ëàíöþгàìè, øâèäê³ñòü г³äðîë³çó ÿêèõ çíèжóºòüñÿ ç³ çìåíøåííÿì ñòóïåíÿ ïîë³ìåð³çàö³¿. Äðóгèé фåðìåíò, ÿêèé áåðå ó÷àñòü â ðîçùåïëåíí³ êñèëàí³â, -åêçî 1,4 β d êñèëîçèäàçà (ÊÔ 3.2.1.37) àáî β êñèëîçèäàçà, ÿêà êàòàë³çóº г³äðîë³ç îë³гîñàõàðèä³â øëÿõîì ïîñë³äîâíîгî â³äùåïëåííÿ çàëèøê³â d êñèëîçè ç íåðåäóêóþ÷îгî ê³íöÿ. Âñòàíîâëåíî, ùî àêòèâí³ñòü êñèëîçèäàçè íàéá³ëüøà ïî â³äíîøåííþ äî êñèëîá³îçè ³ çìåíøóºòüñÿ ç³ çá³ëüøåííÿì äîâжèíè ëàíöþгà ñóáñòðàòà.Äжåðåëàìè êñèëàíàç ìîжóòü áóòè áåçõðåáåòí³ òâàðèíè, âèù³ ðîñëèíè, íàñ³ííÿ, ìîðñüê³ âîäîðîñò³, àëå íàéá³ëüø òåõíîëîг³÷íèìè º ì³êðîîðгàí³çìè -гðèáè, áàêòåð³¿, äð³жäж³ [1]. Ì³êðîáí³ êñèëàíàçè çàâäÿêè ¿õ âèñîê³é ñïåöèф³÷íîñò³, ì'ÿêèì óìîâàì ðåàêö³¿, íåçíà÷íèì âèòðàòàì ñóáñòðàòà ³ óòâîðåííþ ïîá³÷íèõ ïðîäóêò³â, º íàéá³ëüø ïåðñïåêòèâíèìè êàòàë³çàòîðàìè äëÿ г³äðîë³çó êñèëàíó. Àëå âàðò³ñòü фåðìåíòàòèâíîгî г³äðîë³çó á³îìàñè º îäíèì ç фàêòîð³â, ÿêèé ë³ì³òóº åêîíîì³÷íó äîö³ëüí³ñòü ïðîöåñó. Ïðîäóêö³ÿ êñèëàíàç òèì íå ìåíø ìîжå áóòè ïîêðàùåíà çíàõîäжåííÿì á³ëüø ïîòóжíèõ øòàì³â гðèá³â àáî áàêòåð³é, àáî îäåðжàííÿì ìóòàíòíèõ øòàì³â, çäàòíèõ ïðîäóêóâà...

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“…Construction of the recombinant E. coli producing xylanases free of cellulolytic enzymes has been reported [8] . However, very little attention has been paid to its process development which normally related to optimization, kinetic and modeling as well as fermentation modes.…”

Section: Introductionmentioning

confidence: 99%

Kinetics of Xylanase Fermentation by Recombinant <i>Escherichia coli</i> DH5α in Shake Flask Culture

Rusli¹

2009

American Journal of Biochemistry and Biotechnology

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Problem statement: Interest in xylanase enzyme application has markedly increased in pulp and paper processing industries. The switch to xylanase-producing recombinant Escherichia coli DH5α pTP510 is seen here as an economic alternative towards higher productivity and easier downstream purification. Modeling of E. coli DH5α growth and enzyme secretion is thus desired for future optimization in fermentation process. Approach: Kinetics of intracellular xylanase fermentation by a recombinant E. coli DH5α was studied in shake flask culture. The effect of different medium formulations (complex, minimal and defined), initial pH (6.5, 7.0, 7.4 and 8.0) and agitation speeds (150, 200 and 250 rpm) on cell growth and xylanase production were evaluated. Mathematical models based on Logistic and Luedeking-Piret equations had been proposed to describe the microbial growth and xylanase production. Results: Highest xylanase production was obtained in defined medium. Based on medium formulation, the highest cell concentration (4.59 g L −1 ) and xylanase production (2, 122.5 U mL −1 ) was obtained when (NH 4 ) 2 HPO 4 was used as the main nitrogen source, with an adjustment of the initial pH to 7.4 and agitation speed of 200 rpm. The maximum specific growth rate (µ max ), growth associated xylanase production coefficient (α) and non-growth associated xylanase production coefficient (β) was 0.41 h −1 , 474.26 U mg cell −1 and 0 U mg cell −1 h −1 , respectively. Conclusion: Xylanase production was growth associated process and the enzyme secretion was greatly dependent on cell concentration and the specific growth rate of E. coli DH5α.

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Characterization of the recombinant xylanases in Escherichia coli from an alkaliphilic thermophilic Bacillus sp. NCIM 59

Cited by 14 publications

References 20 publications

Isolasi Identifikasi Bakteri Penghasil Xilanase serta Karakterisasi Enzimnya

Isolasi Identifikasi Bakteri Penghasil Xilanase serta Karakterisasi Enzimnya

Xylanases of Microorganisms

Kinetics of Xylanase Fermentation by Recombinant <i>Escherichia coli</i> DH5α in Shake Flask Culture

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