استفاده از سوبسترای آرد عدس قرمز برای تولید آنزیم ناتوکیناز توسط باسیلوس سابتیلیس ناتو

نویسندگان
علی اکبر غلامحسین پور 1
فاطمه میثاق 1
محمدحسین مروت 2
یونس قاسمی 2
1 گروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه جهرم، جهرم، ایران
2 گروه بیوتکنولوژی دارویی، دانشکده داروسازی، دانشگاه علوم پزشکی شیراز، شیراز، ایران
10.22034/FSCT.22.160.173
چکیده
در این پژوهش شرایط تولید بهینه ناتوکیناز با استفاده از باکتری باسیلوس سابتیلیس بر روی سوبسترای عدس دال مورد بررسی قرار گرفت. برای بهینه‌سازی فرایند تخمیر، اثر سه متغیر زمان تخمیر (24، 48 و 72 ساعت)، غلظت ملاس (3، 5 و 7 درصد) و مقدار آب (50، 75 و 100 میلی‌لیتر) بر میزان فعالیت فیبرینولیتیکی و پروتئازی مورد بررسی قرار گرفت. نتایج در قالب طرح مرکب مرکزی (CCD) بررسی و به روش سطح پاسخ (RSM) مدل‌سازی و تجزیه و تحلیل شد. ضریب تبیین مدل‌های رگرسیونی برازش شده برای فعالیت‌های فیبرینولیتیک (مدل درجه دوم) و پروتئازی (مدل خطی) به‌ترتیب 15/97 و 12/90 درصد بوده و فاکتور عدم برازش آنها در سطح اطمینان 95 درصد معنی‌دار نبود، از این رو صحت مدل‌ها برای برازش اطلاعات تایید گردید. با افزایش میزان آب، ملاس و مدت زمان تخمیر، فعالیت فیبرینولیتیک به‌ترتیب افزایش، کاهش و افزایش یافت (p≤0.05). با افزایش مقدار آب، فعالیت پروتئازی نیز افزایش یافت، اما این افزایش معنی‌دار نبود، در حالی‌که افزایش میزان ملاس و مدت زمان تخمیر به‌ترتیب منجر به کاهش و افزایش معنی‌دار فعالیت پروتئازی گردیدند (p≤0.05). در ارتباط با بهینه‌سازی، شرایط بهینه تخمیر عبارت بود از: 38/92 میلی‌لیتر آب، 66/3 درصد ملاس و 90/70 ساعت تخمیر که در چنین شرایطی میزان فعالیت آنزیم‌های فیبرینولیتیک و پروتئاز به‌ترتیب 03/2476 و 68/1 واحد بر گرم پیش‌بینی گردید. برای اعتبارسنجی مدل، تخمیر تحت شرایط بهینه انجام شد و پاسخ‌های به‌دست آمده با پاسخ‌های پیش‌بینی شده توسط مدل مقایسه گردید. نزدیکی داده‌های آزمایشگاهی به مقادیر پیش‌بینی شده توسط مدل صحت مدل را تایید نمود. نتایج این پژوهش نشان داد که از عدس دال می‌توان به‌عنوان سوبسترای تخمیر توسط باکتری باسیلوس سابتیلیس جهت تولید آنزیم‌ ناتوکیناز استفاده نمود.
کلیدواژه‌ها
باسیلوس سابتیلیس ناتو
ناتوکیناز
تخمیر
.RSM

موضوعات

عنوان مقاله English

Utilization of Red Lentil Flour Substrate for the Production of Nattokinase Enzyme by Bacillus subtilis Natto

نویسندگان English

Aliakbar Gholamhosseinpour 1
Fatemeh Misagh 1
Mohammad Hossein Morowvat 2
Younes Ghasemi 2
1 Department of Food Science and Technology, Faculty of Agriculture, Jahrom University, Jahrom, Iran
2 Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
چکیده English

In this research, the optimal production conditions of nattokinase using Bacillus subtilis Natto on red lentil flour substrate were studied. To optimize the fermentation process, the effects of three variables of fermentation time (24, 48 and 72 h), molasses concentration (3, 5 and 7%) and water content (50, 75 and 100 ml) on fibrinolytic and protease activities were investigated. The central composite design (CCD) was employed, and the results were modelled and analyzed using response surface methodology (RSM). Coefficients of determination, R2, of fitted regression models for fibrinolytic (quadratic model) and protease (linear model) activities were 97.15 and 90.12%, respectively, and their lack-of-fit was not significant at 95%. Hence, the models for all the responses were highly adequate. Fibrinolytic activity increased significantly (p≤0.05) with increasing water content and fermentation time, while increasing the amount of molasses decreased fibrinolytic activity (p≤0.05). With increasing the amount of water, protease activity also increased, but this increase was not significant, while increasing the amount of molasses and fermentation time led to a significant decrease and increase in protease activity (p≤0.05), respectively. Concerning optimization, the optimal fermentation conditions were determined as 92.38 ml of water, 3.66% molasses, and 70.90 h of fermentation. Under these conditions, the activities of fibrinolytic enzymes and proteases were predicted to be 2476.03 and 1.68 U/g, respectively. For validation of the model, the optimal sample was produced, and the experimental responses were compared with the responses predicted by the model. The experimental values obtained were quite close to those predicted by the model, indicating the validity of the model. The results of this study showed that red lentil could be used as a substrate for fermentation by B. subtilis to produce the enzyme nattokinase.

کلیدواژه‌ها English

B. subtilis Natto
Nattokinase
Fermentation
RSM
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