160
A. Mukherjee
and candy making, artificial honey), beverage (in soft drinks) and pharmaceutical
industries (Kulshrestha et al. 2013). The most successful commercial invertase is
derived from Baker’s yeast. Though phytoenzymes in food industry offer new means
to address energy savings and increase in efficiency and sustainability, they have not
been seriously commercialized except that of malt amylase (from barley), papain
and bromelain. Production of enzymes from microbial sources requires maintained
upstream process technology, which is cost-effective, but no such technology is
required for extracting phytoenzymes. The source plant itself acts like a bioreactor
producing the enzyme, which needs only to be harvested and extracted. Plant parts
such as stems, producing appreciable amount of extracellular enzymes, are a cheap
source, as these renewable parts can be harvested time to time (Mukherjee et al.
2012).
Mentha spicata L. (Lamiaceae) commonly known as spearmint, (in India as
‘Pudina’), is known to be endowed with a variety of biological properties and is
extensively used in culinary purpose and ayurvedic and folk medicines in the Indian
subcontinent (Güney et al. 2006; Kumar et al. 2008). However, till date, no reports
are available on the presence of macromolecules (enzymes) from the plant. This
is the first report of the presence of appreciable amount of any enzymatic activity
(invertase) in the stems of Mentha spicata.
2 Materials and Methods
2.1 Chemicals
Sucrose, bovine serum albumin (BSA, Fraction-V), DEAE-Sephadex (A50),
Bradford reagent, pepsin, trypsin (porcine pancreas), chmymotrypsin (porcine
pancreas), proteinase-K, maltose, raffinose, inulin (chicory root), melezitose, levan,
N-ethylmaleimide (NEM), p-chloromercuri benozoate (PCMB), iodoacetic acid,
iodoacetamide and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide-hydrochloride
(EDAC) were all purchased from Sigma Chemicals, (St Loius, MO, USA). 3,5dinitrosalicylic acid (DNSA) and 5,5
-dithiobis-2-nitrobenzoic acid (DTNB) were
products of SRL.
2.2 Extraction of Enzyme
Mentha spicata stems (400 g) were chopped into small pieces (1–2 cm long) and
dropped immediately in 400 ml of 0.1 M phosphate buffer, pH 6.0 kept at 37 °C. The
mixture was blended for 5 min in a warring blender and the biomass squeezed through
A. Mukherjee
and candy making, artificial honey), beverage (in soft drinks) and pharmaceutical
industries (Kulshrestha et al. 2013). The most successful commercial invertase is
derived from Baker’s yeast. Though phytoenzymes in food industry offer new means
to address energy savings and increase in efficiency and sustainability, they have not
been seriously commercialized except that of malt amylase (from barley), papain
and bromelain. Production of enzymes from microbial sources requires maintained
upstream process technology, which is cost-effective, but no such technology is
required for extracting phytoenzymes. The source plant itself acts like a bioreactor
producing the enzyme, which needs only to be harvested and extracted. Plant parts
such as stems, producing appreciable amount of extracellular enzymes, are a cheap
source, as these renewable parts can be harvested time to time (Mukherjee et al.
2012).
Mentha spicata L. (Lamiaceae) commonly known as spearmint, (in India as
‘Pudina’), is known to be endowed with a variety of biological properties and is
extensively used in culinary purpose and ayurvedic and folk medicines in the Indian
subcontinent (Güney et al. 2006; Kumar et al. 2008). However, till date, no reports
are available on the presence of macromolecules (enzymes) from the plant. This
is the first report of the presence of appreciable amount of any enzymatic activity
(invertase) in the stems of Mentha spicata.
2 Materials and Methods
2.1 Chemicals
Sucrose, bovine serum albumin (BSA, Fraction-V), DEAE-Sephadex (A50),
Bradford reagent, pepsin, trypsin (porcine pancreas), chmymotrypsin (porcine
pancreas), proteinase-K, maltose, raffinose, inulin (chicory root), melezitose, levan,
N-ethylmaleimide (NEM), p-chloromercuri benozoate (PCMB), iodoacetic acid,
iodoacetamide and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide-hydrochloride
(EDAC) were all purchased from Sigma Chemicals, (St Loius, MO, USA). 3,5dinitrosalicylic acid (DNSA) and 5,5
-dithiobis-2-nitrobenzoic acid (DTNB) were
products of SRL.
2.2 Extraction of Enzyme
Mentha spicata stems (400 g) were chopped into small pieces (1–2 cm long) and
dropped immediately in 400 ml of 0.1 M phosphate buffer, pH 6.0 kept at 37 °C. The
mixture was blended for 5 min in a warring blender and the biomass squeezed through