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The present study describes the production optimization of recombinant L-asparaginase II of Pectobacterium carotovorum MTCC 1428 in Escherichia coli BL21 (DE3) at batch and fed batch bioreactor level. Production of recombinant L-asparaginase II in batch and fed batch mode was found to be 1.34 and 5.38 folds higher, respectively as compared to shake flask culture. SDS-PAGE and native PAGE of the purified enzyme revealed that molecular mass of the subunits and native enzyme are ~37.5 kDa and ~150 kDa, respectively. Optimum range of pH and temperature for hydrolysis of L-asparagine were found to be 7.5 -8.5 and 47˚C -52˚C, respectively. The recombinant enzyme is very specific for its natural substrate, L-asparagine. The activity of recombinant L-asparaginase II is improved by mono cations and diverse effectors including Na + , K + , L-cystine, L-histidine, glutathione and 2-mercaptoethanol whereas, it is moderately inhibited by different divalent cations and thiol group blocking reagent. The kinetic parameters Km, Vmax, kcat and Km/Kcat of purified recombinant L-asparaginase II were determined. The purified L-asparaginase II possesses no partial glutaminase activity, which is prerequisite to reduce the possibility of side effects during the course of anti-cancer therapy.

Background Patient Priorities Care (PPC) aims to identify and integrate patient goals and preferences into health care decision-making to provide more personalized care for multimorbid older individuals. Home-based primary care (HBPC) is a model of care delivery that supports aging in place. HBPC-integrated pharmacists can identify patient priorities and communicate with the team to ensure care is aligned with what matters most. Objectives Evaluate patients’ perceptions of having priorities identification conversations with the pharmacist; identify the value domains represented by patients’ health outcome goals. Setting HBPC program at a large family medicine practice where pharmacists are core members of the interdisciplinary team. Intervention Pharmacists led priorities identification conversations for patients newly enrolled in HBPC. Care preferences and health outcome goals were documented in the medical record and communicated during HBPC team meetings. Design This was a prospective, observational study of HBPC enrollees. After the priorities identification conversation, a three-question survey was administered to identify patients’ perceptions of the conversation and interaction with the pharmacist. Health outcome goals and care preference statements were reviewed to determine with which value domain(s) they most aligned. Descriptive statistics were used for results analysis. Results Pharmacists led conversations with 30 participants. Average overall satisfaction with the conversation was 4.6 on a 5-point Likert scale (1 = least, 5 = most satisfied). Ninety-three percent of patients felt it was appropriate to have a pharmacist lead these conversations. Ninety-seven percent believed it was important/very important to discuss their values and goals with their health care team. The predominant value domains represented were Managing Health (43%) and Functioning (40%). Conclusion Patients were mostly satisfied with having PPC conversations and felt it was appropriate for a pharmacist to lead these conversations. Managing health conditions and preserving function were the most frequent value domains associated with patients’ goals and care preferences.