Journal of Mazandaran University of Medical Sciences
مجله دانشگاه علوم پزشکی مازندران
J Mazandaran Univ Med Sci
Medical Sciences
http://jmums.mazums.ac.ir
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2022
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استخراج و تخلیص لاکتوفرین از شیر شتر و بررسی فعالیت آمیلازی آن
Extraction and Purification of Lactoferrin from Camel Milk and Investigation of Its Amylase Activity
بیوتکنولوژی
Biotechnology
گزارش کوتاه
Brief Report
<span style="font-size:12pt"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="FA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt">سابقه</span></span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt"> و هدف:</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">لاکتوفرین در ترشحات موکوسی، شیر و کلستروم</span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">یافت می</span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.1pt"></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">شود و دارای فعالیت</span></span></span><span lang="AR-SA" style="font-family:"B Zar""><span style="color:black"></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">های ضدمیکروبی، جذب آهن و افزایش پاسخ</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">های ایمنی می</span></span></span><span lang="AR-SA" style="font-family:"B Zar""><span style="color:black"></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">باشد. لاکتوفرین قابلیت تجزیه نشاسته و </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black">ATP</span></span></span><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"> را داراست.</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"> هدف از این مطالعه، بررسی حفظ فعالیت آنزیمی لاکتوفرین شیرشتر پس از خالص سازی توسط کروماتوگرافی</span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">تبادل یونی بود.</span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt">مواد و روش</span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.1pt"></span></span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt">ها:</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">در این تحقیق آزمایشگاهی پس از گرفتن شیر شتر، چربی و کازئین حذف و لاکتوفرین توسط کروماتوگرافی</span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">تبادل یونی با استفاده از </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black">CM-Sephadex-C-50</span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">خالص گردید. مقادیر </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black">K<sub>m</sub></span></span></span><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"> و </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black">V<sub>max</sub></span></span></span> <span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">فعالیت آمیلازی </span></span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">لاکتوفرین، </span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black">pH</span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"> و دمای بهینه واکنش مشخص شد. بدین منظور از محلول 5/0 درصد نشاسته ذرت به</span></span></span></span></span><span lang="AR-SA" style="font-family:"B Zar""><span style="color:black"></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">عنوان سوبسترا و از آزمایش فعالیت یدی برای تعیین مقادیر مصرف نشده استفاده شد. دمای بهینه فعالیت آمیلازی </span></span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">توسط کروماتوگرافی کاغذی تعیین شد.</span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.2pt">یافته</span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"></span></span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.2pt">ها:</span></span></span></span> <a name="_Hlk103544152"><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">نتایج حاصل از بررسی، حضور لاکتوفرین را</span></span></span></span></a> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">در محدوده 73کیلودالتون</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">و</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">غلظت آن را به</span></span></span></span><span lang="AR-SA" style="font-family:"B Zar""><span style="color:black"></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">طور متوسط</span></span></span></span> <span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">µ</span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">g/ml</span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">60 </span></span></span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">نشان داد. </span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">مقادیر </span></span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">K<sub>m</sub></span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"> و </span></span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">V<sub>max</sub></span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><sub> </sub></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">به ترتیب </span></span></span></span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">mg.ml<sup>-1</sup></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">2</span></span></span></span></span></span> <span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">و</span></span></span></span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">mg.ml<sup>-1</sup>.min<sup>-1</sup></span></span></span></span> <span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">0/2 تعیین شد. همچنین لاکتوفرین فعالیت آمیلازی گسترده ای در محدوده </span></span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">pH</span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"> های 4 تا 7 داشت و بیش ترین فعالیت را در ب</span></span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">pH</span></span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"> برابر 5 دارا بود. فعالیت آمیلازی لاکتوفرین در دمای </span></span></span></span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">C</span></span></span></span><span lang="FA" style="line-height:95%"><span style="color:black"><span style="letter-spacing:-.2pt">˚</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">40</span></span></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"> به حداکثر مقدار خود رسید. </span></span></span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">هضم آنزیمی سبب هیدرولیز نشاسته و تبدیل آن به قندهای مالتوز و گلوکز</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt">شد.</span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"></span></span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.2pt">استنتاج:</span></span></span></span> <a name="_Hlk103544162"><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">بر اساس یافتههای مطالعه حاضر روش</span></span></span></span></span></a><span lang="FA" style="font-family:"B Zar""><span style="color:black"> کروماتوگرافی تبادل یونی</span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"> می</span></span></span></span></span><span lang="AR-SA" style="font-family:"B Zar""><span style="color:black"></span></span><span class="StyleComplexZar14pt" style="font-family:"Times New Roman","serif""><span lang="FA" style="font-size:14.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black">تواند سبب جداسازی لاکتوفرین با خلوص بالا گردد.</span></span></span></span></span><span lang="FA" style="font-family:"B Zar""><span style="color:black"> فعالیت آمیلازی بالای لاکتوفرین نشان داد که ساختار پروتئینی آن پس از عبور از ستون کروماتوگرافی حفظ شده است.</span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"></span></span></span></span></span></span></span></span></span></span></span><br>
<strong> </strong><span style="font-size:12pt"><span style="line-height:150%"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif""><span style="letter-spacing:-.2pt">Background and purpose:</span></span></span></span></b> <span style="font-size:10.0pt"><span style="line-height:150%">Lactoferrin is found in mucus, milk, and colostrum secretions and has antimicrobial activities, improves iron absorption, and enhances immune responses. Lactoferrin has the ability to degrade starch. The aim of this study was to investigate the preservation of milk lactoferrin enzymatic activity after purification by ion exchange chromatography.</span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif"">Materials and methods:</span></span></span></b><b> </b><span lang="EN" style="font-size:10.0pt"><span style="line-height:150%">In this experimental research, </span></span><span style="font-size:10.0pt"><span style="line-height:150%">fat and casein were removed from camel milk and lactoferrin was purified by ion exchange chromatography using CM-Sephadex-C-50. K<sub>m</sub> and V<sub>max</sub> values of lactoferrin amylase activity, pH, and optimum reaction temperature were determined. For this purpose, 0.5% solution of cornstarch was used as a substrate and iodine activity test was used to determine the <span style="letter-spacing:-.1pt">unused amounts. The optimum temperature for amylase activity was determined by paper chromatography.</span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif"">Results:</span></span></span></b><b> </b><span style="font-size:10.0pt"><span style="line-height:150%">The presence of lactoferrin was seen at about 73 KD and its average concentration was 60 µg/ml. K<sub>m</sub> and V<sub>max</sub> values were 2mg.ml<sup>-1</sup> and 0.2 mg.ml<sup>-1</sup>.min-1, respectively. Lactoferrin also showed extensive amylase activity in pH 4-7 and had the highest activity at pH 5. The amylase activity of lactoferrin reached its maximum at 40°C. Enzymatic digestion caused hydrolysis of starch and its conversion to maltose and glucose.</span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="vertical-align:top"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif"">Conclusion:</span></span></span></b> <span style="font-size:10.0pt"><span style="line-height:150%">According to the present study, ion exchange chromatography could isolate high purity lactoferrin. High amylase activity of lactoferrin showed that its protein structure was preserved after passing through the chromatographic column.</span></span><i><span style="font-size:10.0pt"><span style="line-height:150%"></span></span></i></span></span></span></span><br>
لاکتوفرین, شیر شتر, کروماتوگرافی تبادل یونی
lactoferrin, camel milk, ion exchange chromatography
175
179
http://jmums.mazums.ac.ir/browse.php?a_code=A-10-14952-1&slc_lang=fa&sid=1
Saeed
Zibaei
سعید
زیبائی
s.zibaee@mrazi.ac.ir
1003194753284600144956
1003194753284600144956
Yes
Associate Professor, Razi Vaccine and Serum Research Institute, Northeast Branch, Department of Research and Development of Biological Products, Agricultural Research, Education and Extension Organization (AREEO), Mashhad, Iran
دانشیار، موسسه تحقیقات واکسن و سرم سازی رازی شعبه شمال شرق، بخش تحقیق و توسعه فرآوردههای بیولوژیک، سازمان تحقیقات، آموزش و ترویج کشاورزی، مشهد، ایران
Anis
Bakhshani
انیس
بخشانی
1003194753284600144957
1003194753284600144957
No
MSc in Cellular and Molecular Biology, Razi University of Kermanshah, Kermanshah, Iran
کارشناسی ارشد بیولوژی سلولی مولکولی، دانشگاه رازی کرمانشاه، کرمانشاه، ایران
Ali
Bidmeshkipour
علی
بید مشکی پور
1003194753284600144958
1003194753284600144958
No
Associate Professor, Department of Biology, Faculty of Science, Razi University of Kermanshah, Kermanshah, Iran
دانشیار، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه رازی کرمانشاه، کرمانشاه، ایران