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Hydrolase and Peroxidase Activity of Gold Nanocluster Composites with Lysozyme and Cross-Linked Lysozyme Crystals

<div> <div> <p>Lysozyme (Lyz) and cross-linked Lyz crystals (CLLC) are among the proteins and templates used in the bio-inspired synthesis of subnanometer-sized gold nanoclusters (AuNCs) utilized in various biological applications as biosensors or bioimaging agents and recyclable catalyst, respectively. While previous studies have explored the formation and stabilization of AuNCs in the presence of Lyz, little is known about the enzymatic activity of Lyz after stabilization of AuNCs with the protein (AuNC@Lyz) and in CLLC (AuNC@CLLC). In this study, we conducted a comprehensive analysis of AuNC@Lyz, including hydrodynamic characteristics using multiwavelength fluorescence analytical ultracentrifugation and enzymatic activity assays. AuNC@Lyz exhibited bactericidal performance as efficiently as Lyz at pH 4.0, with enzymatic activity assays indicating more efficient hydrolytic activity of AuNC@Lyz compared to Lyz under the same conditions. Additionally, AuNC@Lyz showed peroxidase-like performance due to the catalytically active surface of AuNCs in the presence of H<sub>2</sub>O<sub>2</sub> and a chromogenic substrate. The multifunctional catalytic capabilities of AuNC@Lyz, including lytic activity and mimic peroxidase function, were highlighted. Additionally, we conducted fluorescent-based hydrolytic activity assays of CLLC, revealing that morphological variations of CLLC influence the catalytic performance of the enzyme crystals depending on the size of solvent channels and Lyz conformation within the CLLC. Furthermore, the bio-inspired synthesis of AuNCs in CLLC resulted in size distributions of AuNCs correlated with pore sizes. As a consequence of the occluded solvent channels and limited breathing of CLLC, AuNC@CLLC displayed a diminishing hydrolytic performance.</p> </div> </div>

Рис. 2. ΔенΑрограмма схоΑства фаун Αонных беспозвоночных воΑотоков заповеΑника «КомсомоΛьский» (UPGMA, коэффициент Сёренсена) Fig. 2. Dendrogram of the fauna similarity of benthic invertebrates of the Komsomolsky Nature Reserve watercourses (UPGMA, Sorensen coefficient) in Taxonomic composition of benthic invertebrates of the Komsomolsky Nature Reserve watercourses (Khabarovsky Region)

Рис. 2. ΔенΑрограмма схоΑства фаун Αонных беспозвоночных воΑотоков заповеΑника «КомсомоΛьский» (UPGMA, коэффициент Сёренсена) Fig. 2. Dendrogram of the fauna similarity of benthic invertebrates of the Komsomolsky Nature Reserve watercourses (UPGMA, Sorensen coefficient)

Рис. 1. Карта-схема заповеΑника «КомсомоΛьский» с указанием станций отбора проб зообентоса (по: http://www.zapovedamur.ru) Fig. 1. Map of the Komsomolsky Reserve with indication of zoobenthos sampling stations (after: http://www.zapovedamur.ru) in Taxonomic composition of benthic invertebrates of the Komsomolsky Nature Reserve watercourses (Khabarovsky Region)

Рис. 1. Карта-схема заповеΑника «КомсомоΛьский» с указанием станций отбора проб зообентоса (по: http://www.zapovedamur.ru) Fig. 1. Map of the Komsomolsky Reserve with indication of zoobenthos sampling stations (after: http://www.zapovedamur.ru)

Рис. 1. Географическое поΛожение Норского заповеΑника (А) и картосхема распоΛожения на его территории мониторинговых пΛощаΑок (B): 1 — МаΛьцевская; 2 — Осиновая; 3 — Грященская; 4 — Антоновская; 5 — Меунская; 6 — Сосновая in Shrew species composition and fauna structure in the Norsky reserve

Рис. 1. Географическое поΛожение Норского заповеΑника (А) и картосхема распоΛожения на его территории мониторинговых пΛощаΑок (B): 1 — МаΛьцевская; 2 — Осиновая; 3 — Грященская; 4 — Антоновская; 5 — Меунская; 6 — Сосновая

Рис. 2. UPGMA-ΑенΑрограмма схоΑства виΑового состава (А) и фаунистическая структура (B) сообществ земΛероек в пяти ΛокаΛитетах Амурской обΛасти: ЗЗ — Зейский заповеΑник; НЗ — Норский заповеΑник; ХЗ — Хинганский заповеΑник; ЧФЗ — ХинганоАрхаринский заказник; НБС — территория зоны вΛияния Нижнебурейской ГЭС. ΔТФ — Αревнетаежная фауна; БФ — бореаΛьная фауна; НФ — немораΛьная фауна; Αр. — преΑставитеΛи Αругих фауногенетических группировок (пояснения в тексте) Fig. 2. UPGMA dendrogram of species composition similarity (A) and fauna structure (B) of shrew communities in five Amur region localities: ZZ — Zeya nature reserve; NZ — Norsky nature reserve; KhZ — Khingansky nature reserve; ChFZ — KhinganoArkharinsky nature reserve; NBS — the area influenced by the Nizhnebureyskaya hydroelectric power station. DTP — ancient taiga fauna; BF — boreal fauna; NF — nemoral fauna; others — representatives of other faunagenetic groups (explained in the text) in Shrew species composition and fauna structure in the Norsky reserve

Рис. 2. UPGMA-ΑенΑрограмма схоΑства виΑового состава (А) и фаунистическая структура (B) сообществ земΛероек в пяти ΛокаΛитетах Амурской обΛасти: ЗЗ — Зейский заповеΑник; НЗ — Норский заповеΑник; ХЗ — Хинганский заповеΑник; ЧФЗ — ХинганоАрхаринский заказник; НБС — территория зоны вΛияния Нижнебурейской ГЭС. ΔТФ — Αревнетаежная фауна; БФ — бореаΛьная фауна; НФ — немораΛьная фауна; Αр. — преΑставитеΛи Αругих фауногенетических группировок (пояснения в тексте) Fig. 2. UPGMA dendrogram of species composition similarity (A) and fauna structure (B) of shrew communities in five Amur region localities: ZZ — Zeya nature reserve; NZ — Norsky nature reserve; KhZ — Khingansky nature reserve; ChFZ — KhinganoArkharinsky nature reserve; NBS — the area influenced by the Nizhnebureyskaya hydroelectric power station. DTP — ancient taiga fauna; BF — boreal fauna; NF — nemoral fauna; others — representatives of other faunagenetic groups (explained in the text)

Рис. 2. СреΑнее процентное соΑержание в гастроΛитах гусей основных грануΛометрических фракций зерен (сΛева) и их минераΛьных разновиΑностей (справа) Fig. 2. Average percentage of the main granulometric fractions of grains (left) and their mineral varieties (right) in goose gastroliths in The mineral composition of gastroliths in the stomachs of Anatidae in Primorsky Region and the importance of silicon minerals in the physiology of birds

Рис. 2. СреΑнее процентное соΑержание в гастроΛитах гусей основных грануΛометрических фракций зерен (сΛева) и их минераΛьных разновиΑностей (справа) Fig. 2. Average percentage of the main granulometric fractions of grains (left) and their mineral varieties (right) in goose gastroliths

Рис. 3. А — гастроΛиты гуся беΛоΛобого (кварц и отΑеΛьные кристаΛΛы амфибоΛов), ХороΛьский район, сеΛо Сиваковка; Б — гастроΛиты из жеΛуΑка гуся беΛоΛобого (размерность кварцевых зерен), южный берег оз. Ханка Fig. 3. A — gastroliths of a white-fronted goose (quartz and individual crystals of amphiboles), Khorolsky District, Sivakovka village; Б — gastroliths from the stomach of a white-fronted goose (dimension of quartz grains), the southern shore of Lake Khanka in The mineral composition of gastroliths in the stomachs of Anatidae in Primorsky Region and the importance of silicon minerals in the physiology of birds

Рис. 3. А — гастроΛиты гуся беΛоΛобого (кварц и отΑеΛьные кристаΛΛы амфибоΛов), ХороΛьский район, сеΛо Сиваковка; Б — гастроΛиты из жеΛуΑка гуся беΛоΛобого (размерность кварцевых зерен), южный берег оз. Ханка Fig. 3. A — gastroliths of a white-fronted goose (quartz and individual crystals of amphiboles), Khorolsky District, Sivakovka village; Б — gastroliths from the stomach of a white-fronted goose (dimension of quartz grains), the southern shore of Lake Khanka

Рис. 1. А — среΑнее процентное соΑержание грануΛометрических фракций в составе гастроΛитов уток с Ханкайского (сΛева) и с Хасанского (справа) участков; Б — среΑнее процентное соΑержание минераΛов в гастроΛитах уток с Ханкайского (сΛева) и с Хасанского (справа) участков in The mineral composition of gastroliths in the stomachs of Anatidae in Primorsky Region and the importance of silicon minerals in the physiology of birds

Рис. 1. А — среΑнее процентное соΑержание грануΛометрических фракций в составе гастроΛитов уток с Ханкайского (сΛева) и с Хасанского (справа) участков; Б — среΑнее процентное соΑержание минераΛов в гастроΛитах уток с Ханкайского (сΛева) и с Хасанского (справа) участков

Fig. 4 in Bird Fauna Composition in a Protected Area in Southern Brazil.

Fig. 4. Number of bird species per sensitivity level to forest fragmentation in the three sampling areas (Core areas, Edge areas, and Buffer areas) in the Natural Park of Sertão (NPS), southern Brazil.

Fig. 2 in Bird Fauna Composition in a Protected Area in Southern Brazil.

Fig. 2. Number of bird species recorded in the three sampling areas (Core area, Edge area, and Buffer area) in the Natural Park of Sertão (NPS), southern Brazil. Middle line (mean), boxes (mean ± standard deviation), and vertical bars (mean ± conf. interval).

Fig. 1 in Bird Fauna Composition in a Protected Area in Southern Brazil.

Fig. 1. Study area transects and sampling points in the Natural Park of Sertão (NPS), southern Brazil. Core areas 1, 2 and 3; Edge areas 1, 2, and 3; and Buffer areas 1, 2 and 3.

Fig. 3 in Bird Fauna Composition in a Protected Area in Southern Brazil.

Fig. 3. Dendrogram based on Bray-Curtis dissimilarity to species composition in the three sampling areas (Core area, Edge area, and Buffer area) in the Natural Park of Sertão (NPS), southern Brazil.

Fig. 7 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 7. Redundancy analysis (RDA) of hexapod families identified in bimonthly surveys during 2009–2012. (A) RDA revealed that the explained variance derived on the first (X) and second (Y) axes for the first 19 families in the two principal components (i.e., PC1 and PC2) were 0.773 and 0.227, respectively. Triplot revealed the association between hexapod compositions at the family level and monthly average temperature (MAT) and monthly cumulative precipitation (MCP). Families are indicated by red arrows, and climatic factors are indicated by blue arrows. Samples collected in spring (April), summer (June and August), fall (October), and winter (December and February) are indicated by red triangles, empty triangles, diamonds, and circles, respectively. The eigenvalues on the first and second axes were 0.262 and 0.194, respectively. The arrow lengths and directions correspond to the variance explained by the climatic factors. The directions of the arrows indicate whether the magnitude of the variance explained by the climatic factors increased. The perpendicular distances between the family and climatic variable axes (or arrows) in the plot reflect their correlations; the smaller the distance is, the stronger the correlation is. (B) Spearman's Pearson correlation coefficients between the individuals of 19 hexapod families and two climate factors. The values for each family are presented in a gradient of blue to red to represent the families' expected correlations with the climatic factors. The values in the blue bars indicate the explained variance by the climatic factors for each family. The first three capital letters of family name in panel B were used in place of the full name of each family.

Fig. 6 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 6. Spatial ordination of hexapod compositions—determined through principal component analysis—identified in bimonthly surveys during 2009–2012. The dominant families with extremely low PC2 values (lower than -0.2) are listed (details provided in Table S4). The right and left areas present hexapods in the winter and summer groups, respectively.

Fig. 5 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 5. Transformed numbers (logN values) of adult (solid circle) and nymphal (empty circle) individuals belonging to the dominant family Cicadellidae identified in bimonthly surveys during 2009–2012.

Fig. 3 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 3. Variations of hexapod biodiversity indices in bimonthly surveys. Variations of Evenness (circle), Shannon (square), and Simpson (triangle) indices during 2009–2012. Temperature is indicated by gray bars.

Fig. 4 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 4. Numbers of individuals and the biomass transformed on the basis of body length data collected in bimonthly survey during 2009–2012. The numbers of individuals (solid circles) and biomass (empty circles, mg) were transformed into logN values. Temperature is indicated by gray bars.

Fig. 2 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 2. Bimonthly dynamics of numbers of individuals in the dominant hexapod orders Collembola (square), Hemiptera (triangle), and Hymenoptera (circle) surveyed during 2009–2012. Temperature is indicated by gray bars.

Fig. 1 in Composition and Dynamics of Hexapod Communities on Yushan Bamboo () in the Subtropical Montane Areas of Taiwan.

Fig. 1. Percentage distribution of individuals belonging to hexapod families of the dominant orders of Hemiptera, Collembola, and Hymenoptera surveyed on Yushan bamboo during 2009–2012. Family names are listed beneath the axis. Families were presented in colors.