تولید آنزیم لیپاز با استفاده از تکنیک واکسن میکروبی مخلوط از دو جدایه باکتری محلی Kytococcus sedentarius وPseudomonas oleovorans با استفاده از مواد زائد جامد پزشکی

نوع مقاله : مقاله پژوهشی

نویسندگان

گروه زیست شناسی، دانشکده آموزش علوم محض، دانشگاه انبار، انبار، عراق

چکیده

هدف از این مطالعه، جداسازی و شناسایی سویه‌های باکتریایی محلی است که قادر به تجزیه ضایعات پزشکی در حین تولید آنزیم‌های لیپاز هستند. عوامل کلیدی موثر بر تولید آنزیم، از جملهpH، دما و ترکیب مواد مغذی، برای افزایش عملکرد بهینه شدند. این یافته ها به مدیریت پایدار زباله و کاربردهای آنزیم صنعتی کمک می‌کند.
مواد و روش‌ها: این مطالعه در آزمایشگاه‌های گروه علوم زیستی دانشکده آموزش علوم محض دانشگاه انبار انجام شد. جدایه‌های باکتریایی محلی که قادر به تجزیه مواد زائد جامد پزشکی بودند با استفاده از روش‌های تشخیصی استاندارد شناسایی شدند. شناسایی با استفاده از سیستم Vitek تایید شد که جدایه ها را به عنوان Kytococcus sedentarius و Pseudomonas oleovorans طبقه بندی کرد. تولید لیپاز با استفاده از روش کشت غوطه‌ور، با زباله‌های پزشکی جامد به عنوان منبع اصلی کربن و انرژی انجام شد. عوامل مختلفی که بر تولید آنزیم تأثیر می‌گذارند؛ از جمله pH، دما، اندازه تلقیح، منبع نیتروژن، غلظت ضایعات و مدت زمان هچ برای تعیین شرایط بهینه مورد بررسی قرار گرفتند. فعالیت آنزیم اندازه گیری شد و اثر مکمل نیتروژن بر تولید لیپاز ارزیابی شد.
نتایج: جدایه‌های باکتریایی Kytococcus sedentarius و Pseudomonas oleovorans کارایی بالایی در تخریب مواد زائد جامد پزشکی و تولید آنزیم‌های لیپاز نشان دادند. شرایط بهینه برای حداکثر بازده آنزیم به pHبرابر 8، دمای 45 درجه سانتیگراد، حجم تلقیح 6 میلی لیتر در هر 100 میلی لیتر محیط و غلظت ضایعات 2 گرم در هر 100 میلی لیتر محیط تعیین شد. تحت این شرایط، بیشترین فعالیت آنزیمی ثبت شده 5/8 واحد در میلی لیتر بود. افزودن منبع نیتروژن منجر به کاهش فعالیت آنزیم از 5/8 به 1/8 واحد در میلی لیتر شد، که نشان می‌دهد محیط زباله پزشکی جامد نیتروژن کافی را فراهم می‌کند و مکمل‌های اضافی به عنوان بازدارنده تولید آنزیم عمل می‌کنند.
نتیجه‌گیری: یافته‌ها حاکی از آن است که زباله‌های پزشکی جامد می‌توانند به عنوان یک بستر مناسب برای تولید آنزیم عمل کنند و به طور همزمان ضایعات را کاهش دهند و محصولات زیستی ارزشمندی تولید کنند. علاوه بر این، مهار مشاهده شده از فعالیت آنزیم توسط مکمل نیتروژن بر اهمیت بهینه سازی ترکیب مواد مغذی تأکید می‌کند. این تحقیق به مدیریت ضایعات پایدار و تولید آنزیم صنعتی کمک می‌کند و راه را برای مطالعات بیشتر در مورد برنامه‌های کاربردی در مقیاس بزرگ هموار می‌کند.

کلیدواژه‌ها

عنوان مقاله [English]

Production of lipase enzyme using a mixed microbial vaccine technique from two local bacterial isolates, Kytococcus sedentarius and Pseudomonas oleovorans, utilizing solid medical waste

نویسندگان [English]

  • Kamal Saadoon Al-Mufarji
  • Dhafer Fakhri Al-Rawi
Department of Biology, College of Education for Pure Sciences, University of Anbar, Anbar, Iraq
چکیده [English]

Objective
This study aims to isolate and identify local bacterial strains capable of degrading medical waste while producing lipase enzymes. Key factors influencing enzyme production, including pH, temperature, and nutrient composition, were optimized to enhance yield. The findings contribute to sustainable waste management and industrial enzyme applications.
Materials and Methods
This study was conducted in the laboratories of the Department of Life Sciences, College of Education for Pure Sciences, University of Anbar. Local bacterial isolates capable of degrading solid medical waste were obtained and identified using standard diagnostic methods. Identification was confirmed using the Vitek system, which classified the isolates as Kytococcus sedentarius and Pseudomonas oleovorans. Lipase production was carried out using the submerged culture technique, with solid medical waste serving as the primary carbon and energy source. Various factors affecting enzyme production—including pH, temperature, inoculum size, nitrogen source, waste concentration, and incubation duration—were investigated to determine optimal conditions. Enzyme activity was measured, and the effect of nitrogen supplementation on lipase production was assessed.

Results
The bacterial isolates Kytococcus sedentarius and Pseudomonas oleovorans demonstrated high efficiency in degrading solid medical waste and producing lipase enzymes. Optimal conditions for maximum enzyme yield were determined to be pH 8, a temperature of 45°C, an inoculum volume of 6 mL per 100 mL of medium, and a waste concentration of 2 g per 100 mL of medium. Under these conditions, the highest recorded enzyme activity was 8.5 units/mL. The addition of a nitrogen source led to a decrease in enzyme activity from 8.5 to 8.1 units/mL, suggesting that the solid medical waste medium provided sufficient nitrogen, with additional supplementation acting as an inhibitor of enzyme production.
Conclusions
This study successfully isolated and identified two bacterial strains, Kytococcus sedentarius and Pseudomonas oleovorans, with high efficiency in degrading solid medical waste and producing lipase enzymes. The optimal conditions for maximum enzyme production were established, highlighting the potential of these bacteria for biotechnological applications. The findings suggest that solid medical waste can serve as a viable substrate for enzyme production, simultaneously reducing waste and generating valuable bioproducts. Furthermore, the observed inhibition of enzyme activity by nitrogen supplementation underscores the importance of optimizing nutrient composition. This research contributes to sustainable waste management and industrial enzyme production, paving the way for further studies on large-scale applications.

کلیدواژه‌ها [English]

  • factors affecting lipase production
  • Kytococcus sedentarius
  • lipase
  • mixed microbial vaccines
  • Pseudomonas oleovorans

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مجله بیوتکنولوژی کشاورزی
دوره 17، شماره 1
فروردین 1404
صفحه 247-270

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