Papain

In papain-CPC DDM, 2–3 ml of 4% papain-4% CPC working solution [the stock solutions of CPC and papain were prepared by dissolving 4 g of CPC (Himedia, Mumbai, India)] and 4 g of papain (Himedia, Mumbai, India) in 100 ml of distilled water] was added to a 14 ml BD Falcon tube containing 2–3 ml of sputum aliquot. The contents were mixed by vortexing (1 min), and were incubated at 37°C for 30 min. After incubation, the volume was brought to 14 ml with the addition of distilled water. Then, this mixture was thoroughly mixed by inversions and vortexing (1 min) and centrifuged at 3000 x g for 20 min (R-83, Remi, Mumbai, India). The supernatant was carefully decanted and the remaining sediment was re-suspended in 200 μl of distilled water. This re-suspended pellet (0.2 ml) was inoculated on two slants of LJ media (one media with glycerol and the other with sodium pyruvate) followed by incubation at 37°C (aerobically) for 8 weeks aerobically with manual monitoring of once per week for the growth of MTB.

Moth bean seeds (MBS-27 variety) were obtained from Mahatma Phule Krishi Vidyapeeth (MPKV), Rahuri, Maharashtra. Angiotensin-converting enzyme (ACE) was extracted and purified from pig lung tissues according to the protocol described by Abdulazeez Mansurah et al. (19 (link)). Pig lung tissues were brought from a local slaughterhouse (Deonar Abattoir, Mumbai). Pepsin (1,000 U/mg, from porcine), pancreatin 3x (3 NF/USP, from porcine pancreas), trypsin (>2,000 U/g, from bovine pancreas), and papain (from papaya latex) were purchased from HiMedia Laboratories Pvt. Ltd., Mumbai. HHL (N-Hippuryl- His- Leu hydrate) and captopril were purchased from Sigma–Aldrich Chemicals Private Limited, Bangalore, India. All the chemicals used were of analytical grade.