Nova Biologica Reperta
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NBR
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http://nbr.khu.ac.ir
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تولید و تعیین خصوصیت بیوشیمیایی فیتاز گرمادوست جدا شده از باکتری باسیلوس آمیلولیکوفاسینس LOR10
Production and biochemical characterization of a thermostable phytase from Bacillus amyloliquefaciens LOR10
بیوتکنولوژی
Biotechnology
مقاله پژوهشی
Original Article
<span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">فیتاز ارزش تغذیه­ای غذاهای گیاهی را با افزایش هضم پذیری پروتئین و در دسترس سازی مواد معدنی از طریق هیدرولیز فیتات در طی هضم در معده یا در طی پردازش غذا بهبود می­بخشد. منابع میکروبی برای تولید تجاری فیتاز پتانسیل قابل­توجهی دارند. بنابراین، هدف این تحقیق غربالگری و جداسازی باکتری­های مولد فیتاز از چشمه آب گرم با پتانسیل قابل توجه می­باشد. شناسایی بهترین سویه با روش مولکولی </span></span></span><span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">16S rDNA</span></span></span><span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;"> انجام شد. بهینه سازی تولید آنزیم در حضور منابع مختلف کربن، نیتروژن و فسفات انجام گردید. فعالیت و پایداری آنزیم در </span></span></span><span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">pH</span></span></span><span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;"> ها، دماها و یون­های مختلف انجام شد. مقایسه توالی ژن </span></span></span><span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">16S rDNA</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">سویه </span></span></span><span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">LOR10</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">با</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">سایر</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">سویه­ها</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">در</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">بانک</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">ژنی</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">با</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">استفاده</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">از</span></span></span> <span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">Clustal omega</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">میزان</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">98</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">درصد</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">همولوژی</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">با</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">باسیلوس</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">آمیلولیکوفاسینس</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">را</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">نشان</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">داد.</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">نتایج</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">بهینه­سازی</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">محیط</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">نشان</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">داد</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">که</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">گالاکتوز،</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">عصاره</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">مخمر</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">و</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">تری­کلسیم</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">فسفات</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">بعنوان</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">بهترین</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">منبع</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">کربن،</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">نیتروژن</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">و</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">فسفات</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">برای</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">تولید</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">فیتاز</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">می­باشند.</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">دمای</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">بهینه</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">فعالیت</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">در</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">دمای</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">70</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">درجه</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">سانتی­گراد</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">به</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">دست</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">آمد.</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">بهترین پایداری فیتاز نیز در </span></span></span><span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">pH</span></span></span><span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;"> حدود 5 تا 8 به دست آمد. فعالیت فیتازی در حضور کلرید­کلسیم، کلرید­روی و سولفات­منیزیم حدود 4/1، 3/2 و 6/1 برابر به ترتیب افزایش یافت. خاطر نشان می­شود که فعالیت فیتازی در حضور </span></span></span><span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">EDTA</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">و</span></span></span> <span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">SDS</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">حدود</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">30</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">درصد</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">کاهش</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">یافت.</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">این</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">نتایج</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">خاطر</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">نشان</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">می­سازد</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">که</span></span></span> <span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;">فیتاز</span></span></span> <span dir="LTR" style="font-weight:normal;"><span style="font-family:Times New Roman,serif;"><span style="font-size:10.0pt;">LOR10</span></span></span><span style="font-weight:normal;"><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;"> پتانسیل قابل­توجهی برای استفاده­های تجاری از جمله مکمل غذایی را داراست.</span></span></span><span style="font-family:B Nazanin;"><span style="font-size:10.0pt;"></span></span><br />
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Phytase can improve the nutritional value of plant-based foods by enhancing protein digestibility and mineral availability through phytate digestion in the stomach and the food processing industry. Microbial sources are more promising for the production of phytases on a commercial scale. The objectives of this exploration were to screening and isolation of phytase-producing bacteria from hot spring with commercial interest. Molecular identification of the best isolate was achieved by the <em>16S rDNA</em> gene. Optimization of phytase production was prepared in the presence of different phosphate, nitrogen, and carbon sources. Enzyme activity and stability were also explored in the presence of different pHs, temperatures, and ion compounds. Comparing the <em>16S rDNA</em> gene sequence of the isolate LOR10 with those in GenBank using Clustal omega shows 98% sequence homology with <em>Bacillus</em> <em>amyloliquefaciens. </em>Medium optimization studies showed that galactose, yeast extract, and tricalcium phosphate were the best sources of carbon, nitrogen, and phosphate for phytase production, respectively. The optimum temperature activity was also observed to be 70<sup> o</sup>C. Phytase stability was at its optimum in a pH range of 5.0–8.0. Phytase activity increased in the presence of CaCl<sub>2</sub>, ZnCl<sub>2</sub>, and MnSO<sub>4</sub> about 1.4, 2.3 and 1.6 folds, respectively. It could be mentioned that phytase activity decreased by about 30 % in the presence of EDTA and SDS. On the basis of the results, it could be concluded that LOR10 phytase has a great potential for commercial interest as an additive to animal plant-based foods.<br />
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باکتریهای تجزیهکننده فیتات, جداسازی, چشمه آبگرم, شناسایی باکتریایی
bacterial identification, hot spring, isolation, phytate-degrading bacteria
390
399
http://nbr.khu.ac.ir/browse.php?a_code=A-10-215-2&slc_lang=en&sid=1
Arastoo
Badoei-dalfard
ارسطو
بدویی-دلفارد
badoei@uk.ac.ir
10031947532846008676
10031947532846008676
Yes
Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran
گروه زیست شناسی، دانشکده علوم، دانشگاه شهید باهنر کرمان، کرمان، ایران
Maryam
Parhamfar
مریم
پرهامفر
abadoei@gmail.com
10031947532846008677
10031947532846008677
No
Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran
گروه زیست شناسی، دانشکده علوم، دانشگاه شهید باهنر کرمان، کرمان، ایران