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. 2022 Jul 27;27(15):4794.
doi: 10.3390/molecules27154794.

Biochemical Characterization and In Vitro Digestibility of Protein Isolates from Hemp (Cannabis sativa L.) By-Products for Salmonid Feed Applications

Arjun H Banskota  1 , Sean M Tibbetts  1 , Alysson Jones  1 , Roumiana Stefanova  1 , Joerg Behnke  1
Affiliations

Biochemical Characterization and In Vitro Digestibility of Protein Isolates from Hemp (Cannabis sativa L.) By-Products for Salmonid Feed Applications

Arjun H Banskota et al. Molecules. .

Abstract

Hemp (Cannabis sativa L.) processing by-products (hemp cake and hemp seed hulls) were studied for their protein content, extraction of protein isolates (PIs), and their in vitro protein digestibility (IVPD). Crude protein contents of hemp cake and hemp seed hulls were 30.4% and 8.6%, respectively, calculated based on generalized N-to-P conversion factor (N × 5.37). Extraction efficiency of PIs from defatted biomass ranged from 56.0 to 67.7% with alkaline extraction (0.1 M NaOH) followed by isoelectric precipitation (1.0 M HCl). Nitrogen analysis suggested that the total protein contents of PIs extracted using three different alkaline conditions (0.5 M, 0.1 M, and pH 10.0 with NaOH) were >69.7%. The hemp by-product PIs contained all essential amino acids (EAAs) required for fish with leucine, valine, and phenylalanine belonging to the five dominant amino acids. Overall, glutamate was the dominant non-EAA followed by aspartate. Coomassie staining of an SDS-PAGE gel revealed strong presence of the storage protein edestin. High IVPD of >88% was observed for PIs extracted from hemp seeds and by-products when evaluated using a two-phase in vitro gastric/pancreatic protein digestibility assay. PIs extracted from by-products were further tested for their antioxidant activities. The tested PIs showed dose-dependent DPPH radical scavenging activity and possessed strong ORAC values > 650 μM TE/g.

Keywords: Cannabis sativa; amino acids; antioxidant; hemp by-products; hemp cake; hemp hulls; hemp seed; in vitro digestibility; protein isolate; total phenolic.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Crude protein content and extraction yield of protein isolates from defatted hemp seeds and defatted by-products. HSHE—hemp hearts; HSWH—hemp whole seed; HSCA—hemp cake; HSHU—hemp seed hulls.
Figure 2
Figure 2
Protein isolate extracted from hemp seeds and hemp by-products. Clockwise from top left: protein isolate from hemp hearts, hemp whole seed, hemp hulls, and hemp cake.
Figure 3
Figure 3
Non-reducing (A) and reducing (B) SDS-PAGE profiles for protein isolates extracted from hemp seeds and hemp by-products at 0.1 M NaOH. HSHE—hemp hearts; HSWH—hemp whole seed; HSCA—hemp cake; HSHU—hemp seed hulls. * Based on previously published data from Wang et al. (2019) [20] and Pavlovic et al. (2019) [21].
Figure 4
Figure 4
Two-phase in vitro gastric/pancreatic protein digestibility (IVPD) of protein isolates (PIs) extracted from hemp seed and by-products. Data are expressed as the means ± SD (n = 4). HSHE—hemp hearts; HSWH—hemp whole seed; HSCA—hemp cake; HSHU—hemp seed hulls.
Figure 5
Figure 5
DPPH radical scavenging activity of protein isolates extracted from hemp seed and by-products. Data are expressed as the means ± SD (n = 3). HSHE—hemp hearts; HSWH—hemp whole seed; HSCA—hemp cake; HSHU—hemp seed hulls.
Figure 6
Figure 6
ORAC values and total phenolic (TP) content of protein isolates extracted from hemp seed and by-products. Data are expressed as the means ± SD (n = 4 for ORAC and n = 3 for TP). HSHE—hemp hearts; HSWH—hemp whole seed; HSCA—hemp cake; HSHU—hemp seed hulls.
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